Nitrosourea derivatives, process for their preparation and medicaments containing them

ABSTRACT

The invention relates to nitrosourea derivatives, a process for their preparation and pharmaceutical compositions containing them. 
     The nitrosourea derivatives of the invention correspond to the following formula (I): ##STR1## in which: R can represent a hydrogen atom or an alkyl group from 1 to 30 carbon atoms, X represents a hydroxy group or an --NR 1  R 2  group, Y represents a hydrogen atom, a hydroxy group or an --NR&#39; 1  R&#39; 2  group, where R 1  and/or R&#39; 1  each represent a hydrogen atom or a ##STR2## group, Hal being a halogen, and R 2  and/or R&#39; 2  can each represent a hydrogen atom or an alkyl group comprising from 1 to 6 carbon atoms, R&#39; and R&#34; can represent hydrogen or OH, provided that at least X represents ##STR3## with R 1  representing ##STR4## or Y represents ##STR5## with R&#39; 1  representing ##STR6## and provided that either R&#39; represents hydrogen or R&#34; represents hydrogen, and R&#39; and R&#34; cannot be simultaneously hydrogen atoms. 
     These compounds present an antitumor activity. No drawing.

CROSS-REFERENCE TO RELATED APPLICATIONS

This is a continuation-in-part of Ser. No. 732,007, filed April 24,1985, now abandoned.

BACKGROUND OF THE INVENTION

The present invention relates to novel nitrosourea derivatives and, morespecifically, to novel 2-desoxy-sugar-nitrosoureas and4-desoxy-sugar-nitrosoureas, to processes for their preparation and totheir therapeutic uses.

It is known that various nitrosoureas have powerful cytostatic andoncostatic activity detected within the framework of pharmacologicalexperiments and clinical treatment: this is the case, in particular, of(1,3-bis-2-chloroethyl)-1-nitrosourea [BCNU] marketed under thetrademark "BICNU" (cf. Dictionnaire VIDAL 1984), of1-(2-chloro-ethyl)-3-cyclohexyl nitrosourea [CCNU] marketed under thetrademark "BELUSTINE" (cf. Dictionnaire VIDAL 1984) and1-(2-chloro-ethyl)-3-(4-methyl-cyclohexyl)nitrosourea [Me CCNU]: cf. G.MATHE and Y. KENIS: Expansion Scientifique, 1975, 3rd Ed. LaChimiotherapie des cancers "(leucemies, hematosarcomes et tumeurssolides)" and T. H. WASSERMAN, M. SLAVIK & S. K. CARTER, Cancer Treat.Rev., 1974, 1, p. 131, "Review of CCNU in clinical cancer therapy". G.P. WHEELER et al (Cancer Res., 1974, 34, 194) attribute their oncostaticaction to an alkylation and a carbomoylation of proteins. It has alsobeen suggested that their lipophilic character is essential in so far asit conditions the passage through cell membranes in particular theblood-brain barrier. However, these compounds have the drawback ofshowing certain toxicity, particularly hematological, at the doses atwhich they are revealed to be active. Consequently, this toxicity limitstheir use at doses less than those which seem necessary for the removalof cancer cells and has incited a team of researchers to aim atobtaining medicaments more active and less toxic than the precedingones, by synthesizing derivatives of nitrosoureas whose hydrophiliccharacter is increased with respect to the preceding ones, such assugar-nitrosoureas, in which the sugar molecule is ribose, xylose orglucose: cf. J. L. IMBACH et al, Biomedicine, 1975, 23, p. 410-413, "Theoncostatic and immunosuppressive action of new nitrosourea derivativescontaining sugar radicals". Thus these authors have established theoncostatic action of the following four compounds on L 1210 leucemia andtheir low toxicity:1-(2-chloro-ethyl)3-(ribofuranosyl-2',3'-isopropylidene-5-paranitrobenzoate)-nitrosourea [RFCNU],3-(2-chloro-1-ethyl-2'-desoxy-glucopyranosyl-14,3',4",6'-tetracetate)nitrosourea[GCNU], 1-(2-chloro-ethyl)3-(ribopyranosyl-2',3',4'-triacetate)-nitrosourea [RPCNU] and1-(2-chloro-ethyl) 3-(xylopyranosyl 2',3',4'-triacetate)-nitrosourea[XPCNU]. These compounds are prepared by reacting the appropriateamino-sugar with 2-chloro-ethyl isocyanate, then by proceding withnitrosation of the urea obtained.

These compounds are in oily form, a difficult physical state to handlein therapeutics. This is why, within the scope of the experimentscarried out on these products, recourse has preferably been had to theirsolidification by blocking the hydroxy groups.

In a subsequent work J. L. MONTERO et al (Eur. J. Med. Chem. ChimicaTherapeutica, mars/avril 1976, 11, n 2, p. 183-187: "Synthese denouvelles glycosylnitrosourees a visees oncostatiques - les1-nitrosoureido-1-desoxy-glucopyranoses"), describedglycosyl-nitrosoureas in which the sugar-nitrogen bond is located in ananomeric position, which has oncostatic activity, namely1-[3-(2-chloro-ethyl) 3-nitroso-ureido]-1-beta-D-glucopyranose and2,3,4,6-tetra-O-acetyl1-[3-(2-chlor-ethyl)nitroso-3-ureido]-1-desoxy-beta-D-glucopyranose, andwhich represents in addition the advantage of lower toxicity on the bonemarrow and not being diabetogenic, whereas streptozotocine or2-desoxy-2-(3-methyl-2-nitrosoureido)-D-glucopyranose, which is acompound of natural origin, presents antibiotic, antineoplasicproperties, and also presents undesirable diabetogenic properties aswell as a high renal and hematological toxicity (cf. Drugs of thefuture, vol. IV, n 2, 1979, p. 137-139).

One of the aspects of the invention is to provide novel nitrosoureaderivatives, for which the profile of the activity curve shows itsmaximum at a dosage far below the threshold of the toxicity.

Another aspect of the invention is to provide novel nitrosoureaderivatives having a good therapeutic index.

It is another aspect of the invention to provide also novel nitrosoureaderivatives having physical properties enabling their use intherapeutics.

Another aspect of the invention is to provide novel nitrosoureaderivatives having a solid and stable form.

SUMMARY OF THE INVENTION

According to the invention these various aspects are achieved by meansof a novel series of derivatives of 2-desoxy-sugar-nitrosoureas and4-desoxy-sugar-nitrosoureas, which are distinguished from knownnitrosourea derivatives, particularly by the nature of the osidesynthon, which is a 2-desoxy-sugar or a 4-desoxy-sugar.

It has been observed that by resorting to 2-desoxy-sugar or a4-desoxy-sugar, substituted on the 3 and/or at the 6 carbon of theabove-said sugar, by a nitrosourea group, and which can carry differentsubstituents on the carbon at the 4 position of the above-said sugar,when it relates to a 2-desoxy-sugar or on the carbon at the 2 positionof the above-said sugar, when it relates to a 4-desoxy-sugar, novelnitrosourea derivatives are obtained whose activity is considerablyincreased and whose toxicity is low with respect to the compoundsalready known. One of the interests of these derivatives can beconnected with the hypothesis according to which the 2-desoxy-sugars andthe 4-desoxy-sugars have a certain lability at the level of the osidelinkage, which can result in the formation of a free sugar in certainbiological media and permit the provision of compounds firstlylipophilic, which then become hydrophilic, which would make easier thepassage of cellular barriers and would account for an increase ofantitumor activity.

An object of the invention is to provide novel nitrosourea derivatives,characterised in that they correspond to the following general formula(I): ##STR7## in which: R represents a hydrogen atom, an alkyl groupfrom 1 to 30, preferably 1 to 12 carbon atoms or an aralkyl group from 7to 12, preferably 7 to 9 carbon atoms, optionally substituted by one orseveral, particularly up to 3, halogen atoms, NO₂, NH₂, CF₃ groups oralkoxy groups of 1 to 4 carbon atoms,

X represents a hydroxy group or an NR₁ R₂ group

Y represents a hydrogen atom a hydroxy group or an ##STR8## group whereR₁ and/or R'₁ each represent a hydrogen atom or a ##STR9## group, Halbeing a halogen, preferably Cl, and R₂ and/or R'₂ each represent ahydrogen atom, an alkyl group comprising from 1 to 6 carbon atoms, anaralkyl group comprising 7 to 12, preferably 7 to 9 carbon atoms, acycloalkyl group comprising from 3 to 6 carbon atoms, an aryl group of 4to 10 carbon atoms, the aryl and aralkyl groups being possiblysubstituted by one or several, particularly up to 3, halogen atoms,NO₂,NH₂,CF₃ groups or alkoxy groups of 1 to 4 carbon atoms,

R' and R" represent hydrogen, OM, M representing an alkyl groupcomprising from 1 to 30, preferably from 1 to 12 carbon atoms, an arylgroup from 4 to 10 carbon atoms, an aralkyl group comprising from 7 to12, preferably from 7 to 9 carbon atoms, the aryl and aralkyl groupsbeing possibly substituted by 1 or several, particularly up to 3,halogen atoms, NO₂,NH₂,CF₃ groups or alkoxy groups from 1 to 4 carbonatoms, or M representing an acyl group from 2 to 8 carbon atoms,preferably 2 or 3, or an aroylgroup from 5 to 12, preferably 5 to 9carbon atoms, unsubstituted or substituted by one or several,particularly up to 3, NO₂, NH₂, CF₃ groups, halogen, alkoxy of 1 to 4carbon atoms, provided that either R' or R" represents hydrogen, R' andR" being not simultaneously hydrogen and provided that at least Xrepresents ##STR10## with R₁ representing ##STR11## or Y represents##STR12## with R'₁ representing ##STR13##

In a preferred class of compounds of the invention, R' represents H andR" represents OH.

In another preferred class of compounds of the invention R" represents Hand R' represents OH.

In Formula I and certain of the Formulas following, linkages have beenshown between the groups R, R', R", CH₂ Y and X on the one hand and thering structure on the other hand, by the symbol . This representationmeans that each of the R,R',R",CH₂ Y and X groups can be either at the αposition or at the β position, according to the HAYWORTH representation,and in an arrangement compatible with the stereochemical requirements.

In the rest of the description, the term alkyl includes linear, branchedor cyclic alkyl groups (cycloalkyl).

A preferred class of compounds according to the invention is constitutedby the compounds corresponding to the Formula II below: ##STR14## inwhich R represents a hydrogen atom, an alkyl group from 1 to 30,preferably 1 to 12 carbon atoms, or an aralkyl group from 7 to 12,preferably 7 to 9 carbon atoms, optionnally substituted by one orseverals particularly up to 3 halogen atoms, NO₂,NH₂,CF₃ groups oralkoxy groups from 1 to 4 carbon atoms.

X represents a hydroxy group or an NR₁ R₂ group

Y represents a hydrogen atom, a hydroxy group or an ##STR15## groupwhere R₁ and/or R'₁ each represent a hydrogen atom or a ##STR16## group,Hal being a halogen, preferably Cl, and R₂ and/or R'₂ represent ahydrogen atom, an alkyl group comprising from 1 to 6 carbon atoms, anaralkyl group comprising from 7 to 12, preferably 7 to 9 carbon atoms, acycloalkyl group comprising from 3 to 6 carbon atoms, an aryl group from4 to 10 carbon atoms, the aryl and aralkyl groups being optionallysubstituted by one or several, particularly up to 3 halogen atoms, NO₂,NH₂, CF₃ group or alkoxy groups of 1 to 4 carbon atoms,

R" represents preferably OH, but can be replaced by OM, M representingan alkyl group comprising from 1 to 30, preferably 1 to 12 carbon atoms,an aryl group from 4 to 10 carbon atoms, an aralkyl group comprisingfrom 7 to 12, preferably 7 to 9 carbon atoms, the aryl and aralkylgroups being optionally substituted by one or several, particularly upto 3 halogen atoms, NO₂, NH₂, CF₃ groups or alkoxy groups of 1 to 4carbon atoms, or M representing an acyl group from 2 to 8 carbon atoms,preferably 2 or 3, or an aroyl group from 5 to 12, preferably 5 to 9carbon atoms, unsubstituted or substituted by one or several,particularly up to 3, NO₂, NH₂, CF₃ groups, halogen, alkoxy from 1 to 4carbon atoms, provided that at least X represents ##STR17## with R₁representing ##STR18## or Y represents ##STR19## with R'₁ representing##STR20##

These compounds of formula (II) represent the particular case of formula(I) in which R' represents hydrogen.

Among the compounds of formula (II), a preferred class of compoundsaccording to the invention is constituted by those of formula (III)below: ##STR21## in which R, R", X and Y have the above-indicatedmeanings.

The compounds according to the invention of Formula III belong to theclass of 4-desoxy,alpha-D-xylohexopyranoside compounds.

Among the compounds of Formula (II), a preferred class of compoundsaccording to the invention is constituted by those of Formula IV below:##STR22## in which R, R", X and Y have the previously indicatedmeanings.

The compounds of Formula (IV) belong to the clas class of4-desoxy,alpha-L-xylohexopyranoside compounds.

A particularly preferred class of compounds according to the inventionis constituted by the compounds corresponding to the Formula V below:##STR23## in which R represents a hydrogen atom, an alkyl group from 1to 30, preferably 1 to 12 carbon atoms or an aralkyl group from 7 to 12,preferably 7 to 9 carbon atoms, optionally substituted by one or severalparticularly up to 3, halogen atoms, NO₂,NH₂,CF₃ groups or alkoxy groupsof 1 to 4 carbon atoms.

X represents a hydroxy group or an NR₁ R₂ group

Y represents a hydrogen atom, a hydroxy group or an ##STR24## groupwhere R₁ and/or R'₁ each represent a hydrogen atom or a ##STR25## group,Hal being halogen, preferably Cl, and R₂ and/or R'₂ each represent ahydrogen atom, an alkyl group comprising 1 to 6 carbon atoms, an aralkylgroup comprising 7 to 12, preferably 7 to 9 carbon atoms, a cycloalkylgroup comprising from 3 to 6 carbon atoms, an aryl group of 4 to 10carbon atoms, the aryl and aralkyl groups being optionally substitutedby one or several, particularly up to 3, halogen atoms, NO₂,NH₂,CF₃groups or alkoxy groups of 1 to 4 carbon atoms

R' represents preferably OH, but OH can be replaced by OM, Mrepresenting an alkyl group comprising from 1 to 30, preferably from 1to 12 carbon atoms, an aryl group of 4 to 10 carbon atoms, and aralkylgroup comprising 7 to 12, preferably 7 to 9 carbon atoms, the aryl andaralkyl groups being optionally substituted by one or several,particularly up to 3, halogen atoms, NO₂,NH₂,CF₃ groups or alkoxy groupsfrom 1 to 4 carbon atoms, or M representing an acyl group of 2 to 8carbon atoms, preferably 2 or 3, or an aroyl group from 5 to 12,preferably 5 to 9 carbon atoms, unsubstituted or substituted by one orseveral, particularly up to 3, NO₂,NH₂,CF₃ groups, halogen, alkoxy of 1to 4 carbon atoms provided that at least X represents ##STR26## with R₁representing ##STR27## or Y represents ##STR28## with R'₁ representing##STR29##

These compounds of Formula V represent the particular case of Formula Iin which R" represents a hydrogen atom.

Among the compounds of Formula V, a preferred class of compounds of theinvention is constituted by those of Formula (VI) below: ##STR30## inwhich R, R', X and Y have the previously indicated meanings.

The compounds of formula (VI) belong to the class of 2-desoxy,alpha-D-arabinohexopyranoside compounds.

Among the compounds of formula (V), another preferred class of compoundsprovided by the invention is constituted by those of the followingformula (VII): ##STR31## in which R, R', X and Y have the previouslyindicated meanings.

These compounds belong to the class of 2-desoxy,alpha-L-arabinohexopyranoside compounds.

A preferred class of compounds according to the invention is constitutedby those of formulad (I), (II), (III), (IV), (V), (VI), and (VII) inwhich:

R represents an alkyl group from 1 to 12 carbon atoms, aralkyl from 7 to12 carbon atoms;

R' or R" represents an OM group, M being an alkyl group comprising from1 to 12 carbon atoms, an aryl group comprising 4 to 10 carbon atoms;

X represents an NR₁ R₂ group, R₁ representing ##STR32## Hal being ahalogen, particularly Cl;

Y represents a hydrogen atom or a hydroxy group.

Another preferred class of compounds according to the invention isconstituted by those of formulae (I), (II), (III), (IV), (V), (VI), and(VII) in which:

R represents an alkyl group from 1 to 12 carbon atoms, halogenoaralkylfrom 4 to 10 carbon atoms;

R' or R" represents an OM group, M being an acyl group of 2 to 8 carbonatoms, an aroyl group from 5 to 12 carbon atoms;

X represents an NR₁ R₂ group, R₁ representing ##STR33## Hal being ahalogen, particularly Cl,

Y represents a hydrogen atom or a hydroxy group.

Another preferred class of compounds according to the invention isconstituted by those of Formulas (I), (II), (III), (IV), (V), (VI), and(VII) and in which:

R represents an alkyl group from 1 to 12 carbon atoms, an aralkyl groupfrom 7 to 12 carbon atoms;

R' or R" represents OH;

X represents an NR₁ R₂ group, R₁ representing ##STR34## Hal representinga halogen, particularly Cl,

Y represents a hydrogen atom.

Another class of preferred compounds according to the invention isconstituted by those of Formula (I), (II), (III), (IV), (V), (VI) and(VII) in which:

R represents an alkyl group from 1 to 12 carbon atoms, an aralkyl groupfrom 7 to 12 carbon atoms;

R' or R" represent OH;

X represents an alkyl amino group, in which the alkyl group has 1 to 6carbon atoms, or arylamino in which the aryl group has 4 to 10 carbonatoms, and

Y represents NR'₁ NR'₂, R'₁ representing ##STR35## Hal representing ahalogen, particularly Cl.

Another class of preferred compounds according to the invention isconstituted by those of Formula (I), (II), (III), (IV), (V), (VI) and(VII) in which:

R represents an alkyl group of 1 to 12 carbon atoms, an aralkyl group of7 to 12 carbon atoms;

R' or R" represents OH;

X represents a hydroxy group,

Y represents NR'₁ R'₂, R'₁ representing ##STR36## Hal representinghalogen, particularly Cl.

Another class of preferred compounds according to the invention isconstituted by those of Formula (I), (II), (III), (IV), (V), (VI) and(VII) in which:

R represents an alkyl group of 1 to 12 carbon atoms, an aralkyl group of7 to 12 carbon atoms,

R' or R" represent OH,

X represents an NR₁ R₂ group, R₁ representing ##STR37## Hal being ahalogen, particularly Cl,

Y represents an alkylamino group, in which the alkyl group has 1 to 6carbon atoms or arylamino in which the aryl group has 4 to 10 carbonatoms.

Another preferred class of compounds according to the invention isconstituted by those of Formula (V), (VI) and (VII) in which:

R represents an alkyl group from 1 to 12 carbon atoms, aralkyl from 7 to12 carbon atoms;

R' represents an OM group, M being an alkyl group comprising 1 to 12carbon atoms, an aryl group comprising 4 to 10 carbon atoms;

X represents a group NR₁ R₂, R₁ representing ##STR38## Hal being ahalogen, particularly Cl,

Y represents a hydrogen atom or a hydroxy group.

Another preferred class of compounds according to the invention isconstituted by those of Formulas (V), (VI) and (VII) in which:

R represents an alkyl group from 1 to 12 carbon atoms, a halogenoaralkylof 4 to 10 carbon atoms;

R' represents an OM group, M being an acyl group of 2 to 8 carbon atoms,an aroyl group from 5 to 12 carbon atoms;

X represents an NR₁ R₂ group, R₁ representing ##STR39## Hal being ahalogen, particularly Cl,

Y represents a hydrogen atom or a hydroxy group.

Another preferred class of compounds according to the invention isconstituted by the Formulas (V), (VI) and (VII) and in which:

R represents an alkyl group from 1 to 12 carbon atoms, an arylkyl groupfrom 7 to 12 carbon atoms;

R' represents OH;

X represents an --NR₁ R₂ group, R₁ representing ##STR40## Hal being ahalogen, particularly Cl,

Y represents a hydrogen atom.

Another class of preferred compounds according to the invention isconstituted by those of Formulas (V), (VI), and (VII) in which:

R represents an alkyl group from 1 to 12 carbon atoms, an aralkyl groupfrom 7 to 12 carbon atoms;

R' represents OH;

X represents an alkylamino group, in which the alkyl group has 1 to 6carbon atoms, or arylamino in which the aryl group has 4 to 10 carbonatoms and

Y represents --NR'₁ R'₂, R'₁ representing ##STR41## Hal being a halogen,particularly Cl.

Another class of preferred compounds according to the invention isconstituted by those of Formula (V), (VI), and (VII) in which:

R represents an alkyl group of 1 to 12 carbon atoms, an aralkyl group of7 to 12 carbon atoms;

R' represents OH;

X represents a hydroxy group,

Y represents --NR'₁ R'₂, R'₁ representing ##STR42## Hal being a halogen,particularly Cl.

Another class of preferred compounds according to the invention isconstituted by those of Formula (V), (VI) and (VII) in which:

R represents an alkyl group of 1 to 12 carbon atoms, an aralkyl group 7to 12 carbon atoms;

R' represents OH;

X represents an NR₁ R₂ group, R₁ representing ##STR43## Hal being ahalogen atom, particularly Cl,

Y represents an alkylamino group, in which the alkyl group has 1 to 6carbon atoms or arylamino in which the aryl group has 4 to 10 carbonatoms.

In accordance with the invention, the novel derivatives of nitrosoureasof the general Formula I are 2,3,6,-tridesoxyα-D-arabinohexopyranosyl-nitrosoureas,2,3-didesoxy-α-D-arabinohexopyranosyl-nitrosoureas,2,6-didesoxy-α-D-arabinohexopyranosyl-nitrosoureas, 2,3,6-tridesoxyα-L-arabinohexopyranosyl-nitrosoureas,2,3-didesoxy-α-L-arabinohexopyranosyl-nitrosoureas,2,6-didesoxy-α-L-arabinohexopyranosyl-nitrosoureas, 3,4,6-tridesoxyα-D-xylohexopyranosyl-nitrosoureas, 3,4-didesoxyα-D-xylohexopyranosyl-nitrosoureas, 4,6-didesoxyα-D-xylohexopyranosyl-nitrosoureas, 3,4,6-tridesoxyα-L-xylohexopyranosyl-nitrosoureas, 3,4-didesoxyα-L-xylohexopyranosyl-nitrosoureas, 4,6-didesoxyalpha-L-xylohexopyranosyl-nitrosoureas.

A preferred class of compounds according to the invention is constitutedby those of the following formula: ##STR44##

It is also an object of the present invention to provide a process forthe preparation of the novel derivatives of general Formula I accordingto the invention, which consists of reacting, in a first step, an osidegroup of general Formula (I bis) ##STR45## in which R, R' and R" havethe meanings indicated above.

X' represents a hydroxy or --NHR₂ group;

Y' represents a hydrogen atom, a hydroxy group or --NHR'₂

R₂ and R'₂ identical or different, represent independently of oneanother, a hydrogen atom, an alkyl group from 1 to 6 carbon atoms, anaralkyl group from 7 to 12 carbon atoms, aryl from 4 to 10 carbon atoms,cycloalkyl from 3 to 6 carbon atoms, the aryl and aralkyl groups beingoptionally substituted by 1 or several particularly up to 3, halogenatoms, NO₂,NH₂ CF₃ groups or alkoxy groups from 1 to 4 carbon atoms, andin which one at least of the groups X' or Y' represent NHR₂ or NHR'₂ on2-halogeno-ethyl isocyanate to convert the --NHR₂ or NHR'₂ group of thecompound of Formula I bis respectively into ##STR46## Hal being ahalogen atom, particularly chlorine, and in a second step, subjectingthe compound obtained at the end of the first step to nitrosation, bymeans of a nitrite of an alkali metal, preferably sodium nitrite, toconvert the ##STR47## groups respectively into ##STR48## or

The process described above can be illustrated by the following diagramin the case where X' represents NHR₂. ##STR49##

In the case where Y' represents --NHR'₂, the process described above canbe illustrated by the following diagram: ##STR50##

The present invention relates also to a process for preparing novelcompounds of the general formula VI according to the invention, whichconsists of reacting, in a first step, an oside group of the generalformula VI bis: ##STR51## in which: R and R' are such as defined above;

X' represents an --NHR₂ or hydroxy group;

Y' represents a hydrogen atom, a hydroxy group or --NHR'₂

R₂ and R'₂, identical or different, represent independently of oneanother a hydrogen atom, an alkyl group from 1 to 6 carbon atoms, anaralkyl group of 7 to 12 carbon atoms, aryl from 4 to 10 carbon atoms,cycloalkyl from 3 to 6 carbon atoms, the aryl and aralkyl groups beingoptionally substituted by one or several, particularly up to 3, halogenatoms, groups NO₂, NH₂,CF₃ or alkoxy groups from 1 to 4 carbon atoms,and in which one at least of the groups X' and Y' represents --NHR₂ orNHR'₂ On a 2-halogeno-ethyl isocyanate to convert the --NHR₂ or NHR'₂group of the compound of Formula VI bis respectively into ##STR52## Halbeing a halogen atom, particularly chlorine, and in a second step tosubject the compound obtained at the end of the preceding step tonitrosation, by means of an alcali metal nitrite, preferably sodiumnitrite to convert the groups ##STR53## respectively into ##STR54##

The present invention relates also to a process for preparing novelcompounds of the general formula VII according to the invention, whichconsists of reacting, in a first step, an oside group of the generalformula VII bis: ##STR55## in which: R and R' are such as defined above;

X' represents an --NHR₂ or hydroxy group;

Y' represents a hydrogen atom, a hydroxy group or --NHR'₂

R₂ and R'₂, identical or different, represent independently of oneanother a hydrogen atom, an alkyl group from 1 to 6 carbon atoms, anaralkyl from 7 to 12 carbon atoms, aryl from 4 to 10 carbon atoms,cycloalkyl from 3 to 6 carbon atoms, the aryl and aralkyl groups beingoptionally substituted by one or several, particularly up to 3, halogenatoms, groups NO₂, NH₂, CF₃ or alkoxy groups from 1 to 4 carbon atoms,and in which one at least of the groups X' and Y' represents --NHR₂ orNHR'₂ On a 2-halogeno-ethyl isocyanate to convert the --NHR₂ or NHR'₂group of the compound of Formula VII bis respectively into ##STR56## Halbeing a halogen atom, particularly chlorine, and in a second step tosubject the compound obtained at the end of the preceding step tonitrosation, by means of an alcali metal nitrite, preferably sodiumnitrite to convert the groups ##STR57## respectively into ##STR58##

To prepare the compounds of formula (Ibis), which enter into thepreparation of the compounds according to the invention, it is possibleto resort to one of the processes described below.

I Preparation of the compounds of formula ##STR59## IA Firstmodification

The compounds of formula: ##STR60## R, R', R", R₂ and Y' having thepreviously indicated meanings, which enter into the preparation of thecompounds of formula: ##STR61## of the invention, can be obtained fromthe compounds of formula: ##STR62## which are subjected: in a first stepto the action of an aldehyde ##STR63## to convert the amine into animine; in a second step, to the action of a reducing agent, such as ahydride, for example sodium borohydride or sodium cyanoborohydride, toconvert the imine into a secondary amine.

The process which has just been described can also be applied to theobtaining of compounds of formula: ##STR64## in which R, R', R", R'₂, X'have the above indicated meanings; and to obtaining compounds offormula: ##STR65## and in which R₂ =R'₂.

In the latter case, the amounts of aldehyde and reducing agent employedare doubled.

IB Second modification

The compounds of formula: ##STR66## R, R', R", Y' and R₂ having thepreviously indicated meanings which enter into the preparation of thecompounds according to the invention of formula: ##STR67## can also beprepared from the compounds of formula: ##STR68## which are subjected tothe action of an alkyl halogenoformate, of formula ##STR69##particularly an alkyl chloroformate to give the compounds of formula:##STR70## these compounds being then subjected to a reducing agent, forexample aluminium and lithium hydride, to convert the ##STR71## groupinto NHR₂.

This process can also be applied to the obtaining of compounds offormula: ##STR72## as well as to those of formula: ##STR73##

II Preparation of compounds of formula: ##STR74##

The compounds of formula: ##STR75## in which R, R' and R" have the aboveindicated meanings and Y' represents a hydroxy group can be obtainedfrom compounds of formula: ##STR76## which are reduced, particularly bycatalytic hydrogenation, for example hydrogenation in the presence ofpalladized carbon.

The compounds of formula: ##STR77## in which R and R" have the aboveindicated meanings and Y' represents a hydroxy group, can be obtainedfrom compounds of formula: ##STR78## in which one of the hydrogen atomsor --NH₂ groups in the 3 position is protected, particularly by reactingthe previously represented compounds with trifluoroacetic anhydride togive the compounds of formula: ##STR79## which are treated with analcohol ROH, in an acid medium, to give the compounds of formula:##STR80## the protective group --COCF₃ being simultaneously hydrolized.

The compounds of formula: ##STR81## in which R, R' and R" have the aboveindicated meanings and Y' represents a hydrogen atom, can be obtainedfrom compounds of formula: ##STR82## in which R, R' and R" have theabove indicated meanings and Hal represents halogen, particularlybromine, these compounds being reduced, particularly by catalytichydrogenation, for example in the presence of palladized carbon.

The compounds of formula: ##STR83## in which R, R' and R" have the aboveindicated meanings and X' represents a hydroxy group, can be obtainedfrom the compounds of formula: ##STR84## by reduction, particularly bycatalytic hydrogenation, for example in the presence of palladizedcarbon.

Compounds of formula: ##STR85## in which R, R' and R" have the aboveindicated meanings can be obtained from compounds of formula: ##STR86##which are subjected to reduction, particularly catalytic hydrogenation,for example in the presence of palladized carbon.

III Preparation of compounds of formula: ##STR87##

The compounds of formula: ##STR88## in which R, R' and R" have the aboveindicated meanings can be obtained from the compounds of formula:##STR89## which are subjected to the action of acetyl chloride, then toneutralization with ammonia to give the compounds of formula: ##STR90##which are reduced, particularly by catalytic hydrogenation, for examplein the presence of palladized carbon, to give the compounds of formula:##STR91## which can be converted by the action of an alcohol ROH, in anacid medium, into compounds of formula: ##STR92##

The compounds of formula: ##STR93## in which R, R' and R" have the aboveindicated meanings and Y" represents a halogen atom, particularlybromine, can be obtained from compounds of formula: ##STR94## which aresubjected to an alcohol of formula ROH, in an acid medium to give thecompounds of formula: ##STR95## which are then converted into compoundsof the formula: ##STR96## by removal of the benzoyl group, for exampleby means of a base, particularly an alkali alcoholate, such as sodiummethylate.

The compounds obtained above can then be treated with an alkylatingagent, such as M₂ SO₄ or MX, X representing halogen, M representing analkyl group from 1 to 30, preferably 1 to 12 carbon atoms, an aryl groupof 6 to 12 carbon atoms, an aralkyl group from 7 to 12 carbon atoms,preferably 7 to 9 carbon atoms, the aryl and aralkyl groups beingoptionally substituted by one or several, particularly up to 3 halogenatoms, NO₂, NH₂, CF₃ groups or alkoxy from 1 to 4 carbon atoms, orrepresenting an acyl group from 2 to 8 carbon atoms, preferably 2 or 3,or an aroyl group from 5 to 12, preferably 5 to 9 carbon atoms,unsubstituted or substituted by one or several, particularly up to 3NO₂, NH₂, 3 groups halogen, alkoxy from 1 to 4 carbon atoms, in thepresence of a base such as NaOH, to result in the compound of formula:##STR97## with R'=MO, M having the above indicated meaning.

IV Preparation of compounds of formula: ##STR98##

The compounds of the formula: ##STR99## in which R, R' and R" have theabove indicated meanings and X' represents an OH group, can be obtainedfrom the compounds of formula: ##STR100## in which the OH functions atthe 4 and 6 positions are protected, by causing the above describedcompounds to react, for example on acetic anhydride, in the presence ofpyridine to protect the hydroxy group at the 3 position, to give thecompounds of the formula: ##STR101## which, in the presence ofN-halogeno succinimide, preferably N-bromosuccinimide, and bariumcarbonate give the compounds of formula: ##STR102## which are subjectedto an azotide, particularly an alkali metal azotide, such as sodium, inthe presence of dimethylformamide, to give the compounds of the formula:##STR103## which, in the presence of a base, particularly an alkalialcoholate, such as sodium methylate, give: ##STR104## which can besubjected to M₂ SO₄ or MX (X=halogen) in the presence of a base such asNaOH to give the compounds of the formula: ##STR105## with MO=R', Mhaving the above indicated meaning.

The compounds of formula: ##STR106## in which R, R' and R" have theabove indicated meanings, can be obtained from the compounds of theformula: ##STR107## in which Hal represents a halogen atom, particularlybromine, which are subjected to the reaction of an azotide,particularly, of an alkali metal, such as sodium azotide, to give thecompounds of formula: ##STR108## in which, the benzoyl group is removed,for example, by the addition of a base, particularly of an alkalialcoholate such as sodium methylate, to give the compounds of theformula: ##STR109##

These compounds can be converted into compounds of the formula:##STR110## by protecting the hydroxy group at the 4 position,particularly by means of benzyl chloride to obtain compounds of formula:##STR111## then by adding an alcohol of the formula ROH, in an acidmedium, to obtain the compounds of the formula: ##STR112## then byremoving the protective group of the hydroxy function at the 4 position,particularly by means of a base, especially an alkali alcoholate such assodium methylate, to obtain the compounds of the formula: ##STR113##then by alkylating the above compounds, especially by means of M₂ SO₄ orMX, X representing a halogen, M representing an alkyl group from 1 to30, preferably 1 to 12 carbon atoms, an aryl group of 4 to 10 carbonatoms, an aralkyl group from 7 to 12 carbon atoms, preferably 7 to 9carbon atoms, the aryl and aralkyl groups being optionally substitutedby one or several, particularly up to 3 halogen atoms, NO₂, NH₂, CF₃groups or alkoxy of 1 to 4 carbon atoms, or representing an acyl groupof 2 to 8 carbon atoms, preferably 2 or 3, an aroyl group of 5 to 12,preferably 5 to 9 carbon atoms, unsubstituted or substituted by one orseveral, particularly up to 3 NO₂, NH₂, CF₃ groups, halogen, alkoxydefrom 1 to 4 carbon atoms, in the presence of a base such as NaOH, toresult in compounds of the formula: ##STR114## with R'=MO, M having theabove indicated meaning.

The processes which have just been described above are advantageouslyapplied to the preparation of the compounds usable for the synthesis ofcompounds of formula (VI) according to the invention.

More precisely, the process described in the paragraph I, enables thepreparation of compounds of the formula: ##STR115## in which R, R', X',Y', R₂ and R'₂ have the above indicated meanings.

The process described in paragraph II enables the preparation ofcompounds of the formula: ##STR116## in which R, R' and Hal have theabove indicated meanings.

The process described in paragraph III enables the synthesis of thecompounds of the formula: ##STR117## in which R, R' and Hal have theabove indicated meanings.

The process described in paragraph IV enables the preparation ofcompounds of the formula: ##STR118## in which R, R', X' have the aboveindicated meanings.

According to a preferred embodiment of the process according to theinvention, the compounds of the general formula (VIbis) are prepared, toobtain the compounds of formula (VI) according to the invention, byreacting an alpha-D-arabinohexopyranoside of the general formula (VIter)below: ##STR119## in which R and R' are as defined above, X" is anazide, hydroxy, NH₂ group or an alkylamine group, whose alkyl radicalincludes 1 to 6 carbon atoms, Y" can represent halogen, when X"represents an azide or hydroxy group, or Y" represents hydrogen, anazide group, an NH₂ group, hydroxy group or an NHR'₂ group where R'₂ isa hydrogen atom or an alkyl group from 1 to 6 carbon atoms, aralkyl from7 to 12 carbon atoms, aryl from 4 to 10 carbon atoms, or cycloalkyl from3 to 6 carbon atoms, the aryl and aralkyl groups being optionallysubstituted by one or several, particularly up to 3, halogen atoms, NO₂,NH₂, CF₃ groups or alkoxy groups from 1 to 4 carbon atoms,

(a) either with an alcohol in an acid medium, in the case where X" is anazide group and Y" a halogen atom, hydrogen or hydroxy group or in acase where X" is a hydroxy group and Y" is an azide group, R is an alkylgroup from 1 to 12 carbon atoms, aralkyl from 7 to 12 carbon atoms orhalogenoalkyl from 1 to 12 carbon atoms and R' represents OM, M being anacyl group of 2 to 8 carbon atoms or arylester from 6 to 12 carbon atomsand where α-D-arabinohexopyranoside is deoxylated at 2,3,6-, thecompound obtained then being reduced, particularly by catalytichydrogenation, for example in the presence of palladized carbon toconvert the azide group into NH₂,

(b) or with an alkylating agent in the case where X" is an azide group,Y" is a halogen atom or hydrogen, R' is an OH group and R is an alkylgroup from 1 to 12 carbon atoms or aralkyl from 7 to 12 carbon atoms andwhere α-D-arabinohexopyranoside is trideoxylated at 2,3,6-, the compoundobtained being then reduced, particularly by catalytic hydrogenation,for example in the presence of palladized carbon, to convert the azidegroup into an NH₂ group,

(c) or with trifluoroacetic anhydride, in the case where X" is an NH₂group and Y" is a hydroxy group, a hydrogen atom or in the case where X"is a hydroxy group and Y" is an NH₂ group, R is an alkyl group from 1 to12 carbon atoms or aralkyl from 7 to 12 carbon atoms and R' a hydroxygroup, the trifluoro acetamide 3-α-D-arabinohexopyranoside obtainedbeing treated with an alcohol in acid medium to obtain the 3-aminocompound of the corresponding general formula and whereα-D-arabinohexopyranoside is dideoxylated at 2,3, or trideoxylated at2,3,6-;

(d) or with an aldehyde in an alcoholic medium, in the case where X" isan NH₂ group, Y" is a hydrogen atom, an NH₂ group or NHR'₂ group or ahydroxy group or in a case where X" is a hydroxy group and Y" is NH₂ orNHR'₂ group and R and R' are as defined above, to obtain respectivelythe 3-desoxy 3-imino compound and/or the corresponding 6-desoxy 6-iminocompound, which by reduction with a suitable reducing agent such assodium borohydride or cyanoborohydride, gives the amine which thenenables to obtain the α-D-arabinohexopyranoside 3-nitrosourea of thecorresponding general formula VI, 2,3- or 2,6-dideoxylated or2,3,6-trideoxylated.

To prepare the α-D-arabinohexopyranosides-nitrosoureas according to theinvention, procedure is preferably as follows:

Modification (a) of the process:

To a solution of methyl 3-azido 4-O-acyl (or arylester) 6-bromo2,3,6-tridesoxy α-D-arabinohexopyranoside, 0.01 mole, in 200 ml ofhexane, are added 10 ml of alcohol (ethanol, benzyl or other alcohol)and 2 g of paratoluenesulfonic acid.

The solution is heated to reflux for 12 to 48 hours. After cooling, thereaction medium is poured onto a saturated sodium bicarbonate solution,then extracted with ether. The organic phase is evaporated to drynessand gives a crude residue which is chromatographed on silica to give theether of the general formula 1 corresponding to the alcohol.

Modification b of the process:

An alkyl-or aralkyl-3-azido-6-bromo-2,3,6-tridesoxyα-D-arabinohexopyranoside, and preferably methyl-3-azido-6-bromo2,3,6-tridesoxy α-D-arabinohexopyranoside, 0.01 mole, are placed insolution in a suitable solvent such as anhydrous tetrahydrofuran (100ml) or in another volvent such as dioxane, isopropyl, ether, etc . . .5-10 g of NaOH are added, then 5-10 g of an alkylating agent such as analkyl sulfate, an alkyl halide, an aralkyl halide, for example. Thesuspension so-obtained is heated under reflux for 12 to 48 hours. Aftercooling and the slow addition of water, it is stirred at 20° C. for 2hours. The organic phase is withdrawn by decantation, then the aqueousphase is again extracted with 100 ml of tetrahydrofuran. The organicphase, dried over sodium sulfate is evaporated to dryness. 60 to 90% ofthe desired compound are obtained, which is purified by chromotographyon silica.

Modification (c) of the process:

Trifluoroacetic anhydride is reacted with methyl 3-amino 2,3,6-tridesoxyα-D-arabinohexopyranoside prepared by the method of J. BOIVIN et coll.,Carb. Res. 85 (1980) 223-42 to obtain methyl-trifluoro3-acetamido-α-D-arabinohexopyranoside (the methyl group beingreplaceable by another alkyl group or an aralkyl group, as definedabove).

To a solution of 0.01 mole of the trifluoro-3-acetamido derivativeso-obtained, in 200 ml of n-hexane, is added 5 to 20 ml of alcohol and 1to 3 g of p-toluenesulfonic acid. By continuing the treatment asdescribed in the first modication of the process, the ether of thegeneral formula I corresponding to the alcohol is obtained, after havingliberated the protected amine, by means of potassium carbonate, in anaqueous alcoholic medium.

Modification (d) of the process:

An aldehyde is reacted with the corresponding amine (Cf. 3rdmodification of the process) as follows, to obtain the correspondingimine: a solution of 0.01 mole of aldehyde in ethanol or the methanol isadded drop by drop, at ambiant temperature, to a solution of 0.01 moleof the amine which constitutes the starting compound of the thirdmodification of the process, in anhydrous ethanol or methanol. Thesolution is heated under reflux for 2 to 12 hours. After cooling, thereis added, in small portions, from 0.01 to 0.05 ml of a suitable reducingagent such as sodium borohydride or cyanoborohydride, and it is shakenfor 12 hours. After evaporation of the solvent, the residue is taken upagain in 20 ml of water, the precipitate is drained, then dried undervacuum under P₂ O₅. The secondary amine obtained is recrystallized inalcohols.

The process is the same in the case where the 3-amino group is replaced,in the starting compound, by a 3-hydroxy group, the amino being in the 6position. In the case where the desired compound must include asubstitution by a secondary amine not only at the 3-position but also atthe 6-position, the amounts of aldehyde and reducing agent employed aredoubled.

The four modifications of the process described above are illustrated bythe diagrams below:

I--DIAGRAM OF THE MODIFICATION (a) OF THE PROCESS: ETHERIFICATION##STR120## R=Ethyl, benzyl, etc . . . II--DIAGRAM OF THE MODIFICATION(b) OF THE PROCESS ##STR121## and (MO=R' (R and M=methyl, ethyl, benzyl,etc. III--DIAGRAM OF THE MODIFICATION (c) OF THE PROCESS WITH BLOCKINGOF THE AMINE AT THE 3 POSITION ##STR122## IV--DIAGRAM OF THEMODIFICATION (d) OF THE PROCESS ##STR123## if Y=NH₂ ##STR124## with R₂=R'₂.

To prepare the alpha-L-arabinohexopyranosides-nitrosoureas according tothe invention, of formula (VII): ##STR125## and in which the nitrosogroup is on the 3 carbon, and R, R', X and Y have the above indicatedmeanings. Procedure is advantageously as follows.

The compounds of the following formula: ##STR126## is treated withO-methylhydroxylamine hydrochloride and sodium acetate to obtain thecompound of the following formula: ##STR127##

By treating the compound obtained previously by diborane and OH⁻ ions,the compound of formula: ##STR128## is obtained.

This compound is then treated with 2-chloro ethyl isocyanate, to givethe compound of formula: ##STR129##

This compound is then dissolved, for example in formic acid, thentreated with sodium nitrite, to give the compounds of the formula:##STR130##

To prepare the alpha-D-xylohexopyranosides-nitrosoureas of the inventionof formula (III), particularly those including a nitrosourea group in 3position, in particular the compound of the following formula:##STR131## it is possible to use the compound of the following formula:##STR132## which in a first step, is treated with 2-chloro ethylisocyanate, to give the corresponding urea, that is to say methyl3-[3-(2-chloro ethyl) ureido]-3,4,6-tridesoxy alpha-D-xylohexopyranosideof the formula: ##STR133##

The compound indicated above, by treatment with sodium nitrite in formicacid results in the corresponding nitrosourea, that is to say methyl3-[3-(2-chloro ethyl) 3-nitroso ureido] 3,4,6-tridesoxyalpha-D-xylohexopyranoside of the formula: ##STR134##

Except as described below, all the compounds which can be used in thepreparation of the compounds of the formula (I), particularly of formula(VI), can be prepared by applying one or other of the reaction sequencesdescribed above, as well as suitable reaction sequences described in theexamples given below, by way of illustration.

The 2-desoxy and 4-desoxy sugar derivatives of nitrosoureas of formula(I), described above exhibit useful therapeutic properties, notablyanti-tumor properties. These nitrosourea derivatives can be made fromthe corresponding oside compounds of formula (Ibis) which can, in turn,be made from the corresponding primary amines. In this regard, a2-desoxy compound of formula (I) of D-configuration can be made from thecorresponding primary amine which can be made from a corresponding azidecomprising the 2-desoxy function with a hydrogen in position 2.Likewise, a 2-desoxy compound of β-L-configuration of formula (I) can bemade from the corresponding primary amine made from a correspondingazide comprising the 2-desoxy function with a hydrogen in position 2.

The 4-desoxy compounds of formula (I) in which Y represents hydrogen canalso be made by equivalent processes starting either from acorresponding azide comprising the 4-desoxy function with a hydrogen inposition 4, for example, for the 4-desoxy compounds ofβ-L-configuration, or from an intermediary compound comprising a primaryamine group, for example, for the 4-desoxy compound ofα-D-configuration.

However, some nitrosoureas compounds of formula (I) or theintermediates, from which they are made, must be made in other ways thanhave been described above. This is particularly the case for thecompounds of formula (I) in which R' represents hydrogen and Yrepresents hydroxy and which are to be made from the correspondingprimary amines or from corresponding azides with a 4-desoxy function andhydrogen in position 4. As a result, certain 4-desoxy compounds offormula (I) with a D-configuration are a special class of newnitrosourea derivatives. Surprisingly, some members of this specialclass of 4-desoxy compounds have high levels of anti-tumor activitiesthat have never been reached in pharmacological tests which areconsidered as particularly significant today. In addition, a new processhereinafter described provides means for making additional new 4-desoxycompounds, particularly 4-halogen and/or 6-acyl, 4-desoxy compoundswhich also possess remarkably high levels of anti-tumor activity.

These new 4-desoxy nitrosourea derivatives are characterized in thatthey correspond to compounds with the formula IA below: ##STR135## inthe form of one of the two anomers, α or β, in which:

R₁ represents an alkyl group of 1 to 12 carbon atoms, or an aralkylgroup of 7 to 12, preferably 7 to 9, carbon atoms, possibly substitutedin the aromatic nucleus by 1 or more, in particular up to 3, halogenatoms, by 1 to 3 NO₂ or CF₃ groups or alkoxy groups containing 1 to 4carbon atoms,

R₄ represents H or Hal, Hal being a Cl or Br atom, especially Cl,

R₆ represents OH or ##STR136## R corresponding to an alkyl group of 1 to6 carbon atoms, an aryl group, unsubstituted or substituted on thearomatic nucleus by 1 or more atoms, in particular up to 3 halogenatoms, by 1 to 3 NO₂ or CF₃ groups or alkoxy groups of 1 to 4 carbonatoms.

Nu represents the group ##STR137## Hal' corresponding to a halogenchosen from among F, Cl, Br and I, identical with, or different from Haland being in particular chlorine.

In formula IA and some of the formulae presented later the bond betweenthe hydrogen atom and OR₁ group at position 1 of the sugar ring isrepresented by the symbol |H, OR₁.

This representation signifies that the OR₁ group can be either in the αposition or in the β position according to the HAYWORTH representation.

In the remainder of the description the term alkyl includes linear,branched or cyclic (cycloalkyl) alkyl groups.

The substances according to the invention represented by formula IA are3,4-dideoxy-α-D-xylohexopyranosides,3,4-dideoxy-β-D-xylohexopyranosides,3,4-dideoxy-α-D-galactohexopyranosides and3,4-dideoxy-β-D-galactohexopyranosides.

An advantageous class of compounds according to the invention isconstituted by the nitrosourea derivatives corresponding to the formulaIA above and in which:

R₁ represents an alkyl group of 1 to 12 carbon atoms,

R₄ represents a halogen atom,

R₆ represents an OH or ##STR138## R having the meanings indicated above.

In this class of nitrosourea derivatives according to the inventionparticularly advantageous compounds of formula IA indicated above arethose in which:

R₁ represents a CH₃ group,

R₄ represents a halogen atom,

R₆ represents OH or ##STR139## R having the meanings indicated above.

In this same class of nitrosourea derivatives according to the inventionparticularly advantageous substances of formula IA indicated above arethose in which, on the one hand:

R₁ represents a CH₃ group,

R₄ represents Cl,

R₆ represents OH or ##STR140## R having the meanings indicated aboveand, on the other:

R₁ represents a CH₃ group,

R₄ represents a halogen,

R₆ represents ##STR141## R having the meanings indicated above.

Another advantageous class of nitrosourea derivatives according to theinvention is constituted by those corresponding to formula IA indicatedabove in which:

R₁ represents a CH₃ group,

R₄ represents a hydrogen atom,

R₆ represents OH.

Another advantageous class of nitrosourea derivatives according to theinvention is constituted by those corresponding to formula IA indicatedabove in which:

R₁ represents a CH₃ group,

R₄ represents a hydrogen atom,

R₆ represents ##STR142## R having the meanings indicated above.

Among the compounds cited above, the most interesting are those havingthe following formulae: ##STR143##

The present invention also relates to a procedure for the preparation ofthe new derivatives corresponding to formula IA ##STR144## in the formof one of the two anomers, α or β, in which:

R₁ represents an alkyl group of 1 to 12 carbon atoms, or an aralkylgroup of 7 to 12, preferably 7 to 9, carbon atoms, possibly substitutedby 1 or more, in particular up to 3, halogens atoms, by 1 to 3 NO₂ orCF₃ groups or alkoxy groups of 1 to 4 carbon atoms,

R₄ represents hydrogen or Hal, Hal being a halogen atom, in particularCl,

R₆ represents OH or ##STR145##

R having the meanings indicated above.

Nu represents the group ##STR146## Hal' being identical with, ordifferent from Hal and representing a halogen atom, in particularchlorine, characterized in that

(a) in a first series of reactions the compound represented by formulaIIA ##STR147## is made to react in the form of a mixture of the α and βanomers with a glycosylating agent constituted by an alcohol R₁ OH, R₁having the meaning indicated above in order to convert the OH group inposition 1 into the OR₁ group. The product thus obtained is treated withan activated derivative of the acid R-COOH in order to convert the OHgroup in position 6 into the ##STR148## group,

* the α and β anomers are separated by a suitable method and

* each of the anomers, α and β, is made to react with a halogenatingagent in order to introduce a halogen atom at position 4, the compoundobtained being present as one of the anomers α and β and corresponds tothe formula IIIA ##STR149## in which R and R₁ have the meaningsindicated in formula IA,

Hal represents a halogen atom,

(b) subsequently, in a second step, the compound represented by formulaIIIA is subjected to a reduction in order to reduce N₃ to NH₂, thisreduction being carried out, in addition, under conditions such that thehalogen atom in position 4 may or may not be hydrogenolyzed, thisreduction being possibly followed by hydrolysis of the ##STR150## groupin position 6 in order to generate an OH group. The compound obtained oncompletion of the second step is present in the form of one of theanomers α or β and corresponds to the formula IVA: ##STR151## in which:R₁, R₄ and R₆ have the meanings indicated in formula IA

(c) in a third series of reactions the anomers, α or β, represented byformula IVA are subjected to reaction with an halogeno ethyl isocyanatein order to convert NH₂ into ##STR152## and the product thus obtained istreated with the nitrite of an alkali metal, in particular sodiumnitrite, in order to convert ##STR153## into ##STR154## in order toobtain the compound represented by formula IA in the form of one of thetwo anomers, α or β.

In the formula IIA and some of the subsequent formulae the bond betweenthe OH and the carbon atom at position 2 of the cyclic structure isrepresented by the symbol ˜. This representation signifies that the OHgroup can be in either the α position or the β position.

As activated derivative of the acid R-COOH, use may be made of the acidderivatives used for the esterification of the primary hydroxyl groups,such as acid halide, preferably chloride, or anhydride.

As halogenating agent sulfuryl chloride or bromide is preferably used.

The starting material corresponds to formula IIA ##STR155## in the formof a mixture of the α and β anomers is described in the literature(particularly by Redlich in Liebigs Ann. Chem. 1981, p. 1215-1222).

A preferred embodiment of the procedure of the invention for thepreparation of the nitrosourea derivatives corresponding to formula IAcomprises:

the reaction of the compound corresponding to formula IIA above in theform of a mixture of the α and β anomers with a glycosylating agentconstituted by R₁ OH, R₁ having the meaning indicated above in order tolead to the formation of the compound corresponding to the formula VA:##STR156##

the introduction of an acyloxy group, of the formula ##STR157## atposition 6, R having the meaning indicated above in order to give riseto the compound with the formula VIA ##STR158##

the separation of the compound VIA into the two anomers, α and β,corresponding to the formula VIIA ##STR159##

the reaction of the anomers, α or β, corresponding to formula VII, withan halogenating agent in order to give rise to the anomers α or β havingthe formula VIIIA ##STR160## then:

* either the reduction of the α or β anomer corresponding to formulaVIIIA thus obtained under conditions such that N₃ is reduced to NH₂without hydrogenolysis of the halogen atom at position 4 to give the αor β anomer corresponding to formula IXA ##STR161## this reductionbeing:

** either followed by the reaction of the α or β anomer corresponding tothe formula IXA with an halogeno ethyl isocyanate in order to convertthe NH₂ into ##STR162## and the action of a nitrite of an alkali metal,in particular sodium nitrite, in order to convert ##STR163## into##STR164## in order to obtain the compounds corresponding to formula IAin which R₄ represents a halogen atom and R₆ represents ##STR165##

** or followed by the hydrolysis of the ##STR166## group present atposition 6 of the α or β anomer corresponding to formula IX and leadingto the formation of an OH group, followed by the reaction of the α or βanomer corresponding to formula IXA with an halogeno ethyl isocyanate inorder to convert the NH₂ into ##STR167## and by the action of thenitrite of an alkali metal, in particular sodium nitrite, in order toconvert ##STR168## into ##STR169## in order to give rise to thecompounds of the formula IA in which: R₄ represents a halogen atom andR₆ represents OH, or * the reduction of the anomers VIIIA underconditions such that the halogen atom at position 4 is replaced by H andthe N₃ group is reduced to NH₂ to give rise to the α or β anomercorresponding to the formula XA ##STR170## this reduction beingfollowed:

** either by the reaction of the α or β anomer corresponding to formulaXA with an halogeno ethyl isocyanate in order to convert the NH₂ into##STR171## and the action of the nitrite of an alkali metal, inparticular sodium nitrite in order to convert ##STR172## into ##STR173##in order to produce the compounds corresponding to formula IA in whichR₄ represents H, R₆ represents ##STR174##

** or by the hydrolysis of the ##STR175## group in position 6 of the αor β anomer corresponding to formula XA thus generating an OH group andleading to the formation of the α or β anomer corresponding to formulaXIA ##STR176## followed by the reaction of the α or β anomercorresponding to formula XIA with an halogeno ethyl isocyanate in orderto convert the NH₂ into ##STR177## and by the action of the nitrite ofan alkali metal, in particular sodium nitrite, in order to convert##STR178## into ##STR179## in order to produce the α or β anomercorresponding to formula IA in which R₄ represents H, and R₆ representsOH.

According to an attractive embodiment of the procedure of the inventionfor the preparation of derivatives corresponding to formula IA, thereduction of the compounds corresponding to formula IIIA such that N₃ isreduced to NH₂ and Hal is replaced by hydrogen is carried out by meansof tributyl tin hydride in the presence of2,2'-azo-bis-isobutyronitrile.

After the glycosylation of the compound corresponding to formula IIA theactivated derivative of the acid RCOOH made to react with the compoundcorresponding to formula IIA in order to introduce a benzoyl group atposition 6 is advantageously benzoyl chloride, used in the presence ofbis tributyl tin oxide.

Another preferred embodiment of the procedure of the invention for thepreparation of nitrosourea derivatives corresponding to formula IA inwhich R₁, R₄, R₆ and Nu have the meanings indicated above comprises:

the glycosylation of the compound corresponding to formula IIA in theform of the mixture of α and β anomers by means of CH₃ OH in order toobtain the compound corresponding to formula XIIA: ##STR180##

the reaction of the compound corresponding to formula XIIA thus obtainedwith benzoyl chloride in the presence of tributyl tin oxide in order toproduce the compound corresponding to formula XIIIA: ##STR181##

the separation of the two α and β anomers starting from the compoundcorresponding to formula XIIIA

the reaction of either the α or β anomer with SO₂ Cl₂ in order to obtainthe α or β anomer corresponding to formula XIVA ##STR182## and

either the catalytic hydrogenation of the α or β anomer corresponding toformula XIA previously explained in order to give rise to the α or βanomer corresponding to formula XV A ##STR183## followed by

.. either the reaction of the α or β anomer corresponding to formula XVAwith an halogeno ethyl isocyanate in order to convert the NH₂ into##STR184## and by the action of the nitrite of an alkali metal, inparticular sodium nitrite, in order to convert ##STR185## into##STR186## to give the α or β anomer corresponding to formula XVIA##STR187##

.. or by the reaction of the α or β anomer corresponding to formula XVAwith a base, in particular an alkali alcoholate in order to hydrolysethe benzoyl group and generate the α or β anomer corresponding toformula XVIIA ##STR188##

followed by the reaction of the α or β anomer corresponding to formulaXVIIA with an halogeno ethyl isocyanate in order to convert the NH₂ into##STR189## and by the action of the nitrite of an alkali metal, inparticular sodium nitrite, in order to convert ##STR190## into##STR191## to give rise to the α or β anomer corresponding to theformula XVIIIA: ##STR192##

or the reduction of the α or β anomer corresponding to formula XIVA bymeans of tributyl tin hydride in the presence of2,2'-azo-bis-isobutyronitrile to give the α or β anomer corresponding tothe formula XIXA: ##STR193## followed

.. either by the reaction of the α or β anomer corresponding to formulaXIXA with an halogeno ethyl isocyanate in order to convert the NH₂ into##STR194## and by the action of the nitrite of an alkali metal, inparticular sodium nitrite, in order to convert ##STR195## into##STR196## to give the α or β anomer corresponding to formula XXA:##STR197##

.. or by the reaction of the alpha or beta anomer corresponding toformula XIXA with a base, in particular an alkali alocoholate, in orderto hydrolyse the benzoyl group and generate the α or β anomerscorresponding to formula XXIA: ##STR198## followed by the reaction ofthe α or β anomer corresponding to formula XXIA with an halogeno ethylisocyanate in order to convert NH₂ into ##STR199## and by the action ofthe nitrite of an alkali metal, in particular sodium nitrite, in orderto convert ##STR200## into ##STR201## to give the α or β anomercorresponding to formula XXIIA: ##STR202##

In carrying out the procedure according to the invention, the compoundscorresponding to formula IIIA ##STR203## in which:

R₁ represents an alkyl group of 1 to 12 carbon atoms, or an aralkylgroup of 7 to 12, and preferably 7 to 9, carbon atoms, possiblysubstituted on the aromatic nucleus by 1 or more, in particular, up to3, halogen atoms, by 1 to 3 NO₂ or CF₃ groups or alkoxy groups of 1 to 4carbon atoms.

R represents an alkyl group of 1 to 6 carbon atoms, an aryl groupunsubstituted or substituted on the aromatic nucleus by 1 or more atoms,in particular up to 3 halogen atoms, NO₂ groups, CF₃ groups or alkoxygroups of 1 to 4 carbon atoms,

Hal represents a halogen atom, in particular chlorine, are new, andexist in the form of either their α or β anomers.

Among the compounds corresponding to formula IIIA, a preferred group ofsubstances is constituted by those corresponding to formula IIIA inwhich:

R₁ represents an alkyl group of 1 to 12 carbon atoms,

R represents ##STR204## A particularly useful new compound correspondsto the following formula ##STR205## in the form of its α and β anomer.

The compounds corresponding to formula IIIA, in particular the twocompounds corresponding to formula IIIAa (α anomer or β anomer), are keyintermediates of the procedure and represent another feature of thepresent invention.

The examples which follow illustrate this invention and in no way limitit.

EXAMPLES EXAMPLE 1 Preparation of benzyl 3-azido 4-O-benzoyl 6-bromo2,3,6-tridesoxy α-D-arabinohexopyranoside by resorting to the firstmodification of the process.

To a solution of methyl 3-azido 4-O-benzoyl 6-bromo 2,3,6-tridesoxyα-D-arabinchexopyranoside (1.5 g, 4.05 mmol.) in 125 ml of n-hexane, areadded 10 ml of benzyl alcohol and 1 g of p-toluenesulfonic acid. Thesuspension is heated to reflux for 20 hours with azeotropic removal ofthe methanol released.

After cooling, the reaction medium is poured into a saturated solutionof sodium bicarbonate then extracted with ether. The excess benzylalcohol is then removed by azeotropic extraction with water, then withtoluene. The crude residue so-obtained (1.8 g) is chromatographed onsilica H with the mixture hexane/methylene chloride, 2:1 as eluant.

1.5 g of benzyl 3-azido 4-O-benzoyl 6-bromo 2,3,6-tridesoxyα-D-arabinohexopyranoside (84%) are isolated.

MP: 77° C. (hexane)--[α]_(D) ²⁰ : +20° (c: 1%; CHCl₃)

IR_(nujol) : ν 2100 cm⁻¹ (N₃) ν 1760, 1260, 1030 cm⁻¹ (ester) ν 1600,1585 cm⁻¹ (aromatic)

In the same manner there is prepared, for example:

Ethyl 3-azido 4-O-benzoyl 6-bromo 2,3,6-tridesoxyα-D-arabinohexopyranoside.

p-chlorobenzyl 3-azido 4-O-benzoyl 6-bromo 2,3,6-tridesoxyα-D-arabinohexopyranoside.

EXAMPLE 2

Use of the second modification of the process. The following compoundsgiven by way of example, are prepared by alkylation of an alkyl oraralkyl-3-azido 6-bromo, 2,3,6-tridesoxy α-D-arabinohexopyranoside.

methyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-ethylα-D-arabinohexopyranoside

methyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-benzylα-D-arabinohexopyranoside

ethyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-ethylα-D-arabinohexopyranoside

ethyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-benzylα-D-arabinohexopyranoside

benzyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-ethylα-D-arabinohexopyranoside

benzyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-benzylα-D-arabinohexopyranoside.

EXAMPLE 3 Use of the third modification of the process.

To a solution of 300 mg (1.16 mmole) methyl 3-fluoroacetamido2,3,6-tridesoxy α-D-arabinohexopyranoside in 100 ml of n-hexane, areadded 5 ml of benzyl alcohol and 300 mg of dry p-toluene sulfonic acid,and the treatment is as described in Example 1.

After chromatography and crystallization in a hexane-acetone mixture,100 mg (30%) of pure product are isolated.

MP: 165° C. - [α] D: +66 (c: 0.5%, CHCl₃).

The amine is then liberated by the action of potassium carbonate in anaqueous methanol medium.

By proceeding in the manner which has just been described, the followingcompounds, given by way of example, are prepared:

benzyl 3-amino 2,3,6-tridesoxy α-D-arabinohexopyranoside

ethyl 3-amino 2,3,6-tridesoxy α-D-arabinohexopyranoside

benzyl 3-amino 2,3-didesoxy α-D-arabinohexopyranoside

ethyl 3-amino 2,3-didesoxy α-D-arabinohexopyranoside.

EXAMPLE 4

Use of the fourth modification of the process. By proceeding inaccordance with the fourth modification of the process it is possible toprepare, among others, the following compounds, given by way of example:

methyl 3-ethylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

methyl 3-benzylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

methyl 3-ethylamino 2,3-didesoxy α-D-arabinohexopyranoside

methyl 3-benzylamino 2,3-didesoxy α-D-arabinohexopyranoside

benzyl 3-ethylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

benzyl 3-benzylamino 2,3,6-tirdesoxy α-D-arabinohexopyranoside

methyl 6-ethylamino 2,6-didesoxy α-D-arabinohexopyranoside

methyl 6-benzylamino 2,6-didesoxy α-D-arabinohexopyranoside

benzyl 6-ethylamino 2,6-didesoxy α-D-arabinohexopyranoside

benzyl 6-benzylamino 2,6-didesoxy α-D-arabinohexopyranoside.

By doubling the amount of aldehyde and of reducing agent employed, thefollowing secondary amines substituted at the 3,6 positions areprepared:

methyl 3,6-diethylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

methyl 3,6-dibenzylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

benzyl 3,6-diethylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside

benzyl 3,6-dibenzylamino 2,3,6-tridesoxy α-D-arabinohexopyranoside.

EXAMPLE 5 Methyl 3-[3-(2-chloro ethyl) ureido] 2,3,6-tridesoxyalpha-D-arabinohexopyranoside--Compound 1

To a solution of 0.8 g (5×10⁻³ mole) of methyl 3-amino 2,3,6-tridesoxyalpha-D-arabinohexopyranoside (prepared according to the method of J.BOIVIN et coll. Carb. Res. 85 (1980) 223-42) in 2 ml of anhydrous DMFare added, drop by drop, at 0° C. and with stirring, 0.4 ml (5.10⁻³mole) of 2-chloro ethyl isocyanate. After 5 hours stirring, the reactionmixture is evaporated to dryness under vacuum. The residue, afterpurification by chromatography on a silicate column, with CHCl₃ : 95,MeOH: 5 eluant, gives a single spot product crystallizing in anhydrousethyl ether. The crystals were drained and then dried. (0.8 g, yield60%).

ANALYSIS C₁₀ H₁₉ Cl N₂ O₄ : 266.5--Calculated % C: 45.0, H: 7.1, N:10.5--Found % C: 44.9, H: 7.0, N: 10.5. MP: 125°-127°

NMR Spectrum Solvent: DMSO.d₆ : 1.14 (d CH₃ -6' J=6 Hz), 1.46 (t(d)H-2'axJ=J'=12 Hz, J"=4 Hz) 1.90 (dd H-2'_(eq) J=12 Hz, J'=4 Hz) 2.78 (tH-4' J=J'=9 Hz) 3.19 (s OCH₃) 3,28 (m CH₂ -4) 3.42 (m H-5') 3.53 (m CH₂-5) 3.61 (m H-3') 4.58 (d H-1' J=3 Hz) 5.99 (d NH J=8 Hz) 6.14 (t NHJ=J'=6 Hz).

EXAMPLE 6 Methyl 3-[3-(2-chloro ethyl) 3-nitrosoureido] 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 2 (IC 81.1183)

1.2 g (4.5.10⁻³ mole) of methyl 3-[3-(2-chloro ethyl) ureido]2,3,6-tridesoxy α-D-arabinohexopyranoside were dissolved in 8 ml offormic acid. To the solution maintained at 0° C., were added in smallportions and with stirring 2.5 g (0.036 mole) of sodium nitrite. After30 minutes, 10 ml of water were added, the stirring was then maintainedfor one hour. The reaction mixture was poured onto 100 ml of ethylacetate and then dried over sodium sulfate and evaporated to drynessunder vacuum. After purification on a chromatograph column, withsilicate support, an eluant CHCl₃ : 9, MeOH: 1, colorless crystals wereobtained (600 mg, Yield 45%).

ANALYSIS C₁₀ H₁₈ Cl N₃ O₅ : 295.71--Calculated % C: 40.6, H: 6.1, N:14.2--Found % C: 40.9, H: 6.0, N: 13.9. MP: 100° C. [α]_(D) ²⁰ : +92.8°(c: 0.5% CHCl₃)--[α]₃₆₅ ²⁰ : 123.6° (c: 0.5% CHCl₃)

NMR Spectrum (DMSO, d₆) 1.15 (d CH₃ -6' J=6 Hz) 1.78 to 1.94 (m CH₂ -2')3.04 (t H-4' J=J'=9 Hz) 3.25 (s OCH₃) 3.51 (m H-5') 3.60 (t CH₂ -4J=J'=6 Hz) 4.10 (m CH₂ -5, H-3') 4.65 (d H-1' J=3 Hz) 8.48 (d NH J=9Hz).

Mass Spectrum: (M+1): 296.

EXAMPLE 7 Methyl 3-azido 2,3-didesoxy α-D-arabinohexopyranoside Compound3

To 7 g (0.024 mole) of methyl 3-azido 4,6-benzylidene2,3-didesoxy-α-D-arabinohexopyranoside prepared according to RICHARDSONA. C., (Carbohyd. Res. 4 (1967), 422-428), in solution in 400 ml ofanhydrous methanol is added drop by drop and with stirring, 8 ml ofacetyl chloride. After 6 hours stirring at ambient temperature, thesolution is neutralized by bubbling gaseous ammonia through it. Afterevaporation under reduced pressure, the residue is taken up again withacetone under reflux.

The acetone solution is concentrated under reduced pressure. By theaddition of hexane, and then by cooling, white crystals are obtainedwhich are drained, washed with hexane, then dried under vacuum. 3.45 g(71%) is obtained.

ANALYSIS C₇ H₁₃ N₃ O₄ : 203.2 MP: 123°-124° C. [α]_(D) ²⁰ : +183° (c:1%, MeOH)

IR: OH 3340, 3280 cm⁻¹ ; N₃ 2100 cm⁻¹

EXAMPLE 8 Methyl 3-amino 2,3-didesoxy α-D-arabinohexopyranoside Compound4

A solution of 5 g (0.0225 mole) of methyl 3-azido 2,3-didesoxyα-D-arbinohexopyranoside in 10 ml of methanol is stirred 12 hours undera hydrogen atmosphere, in the presence of triethylamine (1 ml) and 10%palladized carbon (1 g). The catalyst is removed by filtration. Thefiltrate, evaporated to dryness under reduced pressure, gives 4.35 g ofcompound 4 in the form of a colorless oil.

This compound is crystallized in hydrochloride form.

ANALYSIS C₇ H₁₅ NO₄, HCl: 213.67 MP: 120° C. (dec.) [α]_(D) ²⁰ : +90°(c: 1%, H₂ O).

EXAMPLE 9 Methyl 3-[3-(2-chloro ethyl) 3-nitrosoureido] 2,3-didesoxyα-D-arabinohexopyranoside--Compound 6 (IC 81.1184)

From compound 4, following the operational method already described,methyl 3-[3-(2-chloro ethyl)ureido] 2,3-didesoxyα-D-arabinohexopyranoside, 5 is obtained by the action of 2-chloro ethylisocyanate.

ANALYSIS C₁₀ H₁₉ ClN₂ O₅ : 282--Calculated % C: 42.5, H: 6.7, N: 9.9Found % C: 42.3, H: 6.8, N: 10.0 MP: 125° C. [α]_(D) ²⁰ : +168° (c:0.25%, CHCl₃)

Then by nitrosation with sodium nitrite, in formic acid, Compound 6 isobtained.

ANALYSIS C₁₀ H₁₈ Cl N₃ O₆ : 311.71--Calculated %: C: 38.5, H 5.8, N 13.5Found %: C: 38.4, H: 5.6, N: 13.6. MP: 118° C. [α]_(D) ²⁰ : +96.2° (c:0.5%, CHCl₃)--[α]₃₆₅ ²⁰ : 118.0° (c: 0.5%, CHCl₃)

NMR Spectrum (DMSO.d₆): 1.82 (t(d) H-2'_(ax) J=J'=12 Hz, J": 4 Hz) 1.88(d(d) H-2'eqJ=12 Hz J'=4 Hz) 3.28 (s OCH₃) 3.43 (r H-4' J=J'=9 Hz) 3.35to 3.53 (m H-5', H-3') 3.49 to 3.67 5ab CH₂ -6' J_(gem) =12 Hz) 3.61 (tCH₂ -4 J=J'=6 Hz) 4.10 (m CH₂ -5) 4.73 (d H-1' J=4 Hz) 8.53 (d NH J=9Hz).

Mass Spectrum (chemical ionization): m/e 312 (Cl₃₅, M+1, Basic peak) andm/e 314 (Cl₃₇, 30%); m/e 280 (M+1-32, 30%) and m/e 282 (10%).

EXAMPLE 10 Methyl 3-azido 6-bromo 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 7

10 g (0.026 mole) of methyl 3-azido 4-O-benzoyl 6-bromo 2,3,6-tridesoyα-D-arabinohexopyranoside prepared according to Hanessian, J. Org. Chem.34 (1969)1045-1053, are dissolved in 100 ml of 1M methanolic sodiummethanolate. After 4 hours stirring at 20° C., the solution is thenneutralized by passage over Amberlite IRA 50 resin, form H+. Thefiltrate, evaporated to dryness under vacuum, is purified bychromatography on silica H-60 and gives 6 g (Yield 84%) of Compound 7.

ANALYSIS C₇ H₁₂ Br N₃ O₃ : 266.22 [α]_(D) ²⁰ : +124° (c: 1%, CHCl₃)

IR Spectrum: 3420-3440 cm⁻¹ (OH) 2100 cm⁻¹ (N₃)

EXAMPLE 11 Methyl 3-azido 6-bromo 2,3,6-tridesoxy 4-O-methylα-D-arabinohexopyranoside--Compound 8

5.5 g (0.021 mole) of compound 7 are dissolved in 200 ml of anhydroustetrahydrofuran. 20 g of soda are added and then 20 ml of methylsulfate. The reaction mixture is brought to reflux for 24 hours. Aftercooling, 100 ml of water were added. The organic phase is drawn off bydecantation then the aqueous phase is again extracted with 100 ml oftetrahydrofuran. The organic phase, dried over sodium sulfate, isevaporated to dryness. 5.7 g (Yield 98%) of compound 8 is obtained. Itis purified by chromatography on silica H (eluant hexane-methylenechloride 1:1).

ANALYSIS C₈ H₁₄ BrN₃ O₃ : 280.25 [α]_(D) ²⁰ : +188° (c: 1%, CHCl₃).

IR Spectrum: ν N₃ 2100 cm⁻¹

EXAMPLE 12 Methyl 3-amino 2,3,6-tridesoxy 4-O-methylα-D-arabinohexopyranoside--Compound 9

5.15 g (0.018 mole) of compound 8 in solution in 100 ml of anhydrousethanol and 3 ml of redistilled triethylamine are placed underhydrogenation in the presence of 2 g of palladium on carbon, at ordinarypressure, for 12 hours. After removal of the catalyst, the solution ispassed over an IR 45 ion exchange resin, OH⁻ form. The solutionevaporated to dryness results in 3.5 g of crude product purified bychromatography on silica H, eluant CH₂ Cl₂ -MeOH 9:1

ANALYSIS C₈ H₁₇ NO₃ : 175.22 [α]_(D) ²⁰ : +109° (c: 1%, CHCl₃)

EXAMPLE 13 Methyl 3-[3-(2-chloro ethyl) 3-nitroso ureido]2,3,6-tridesoxy 4-O-methyl alpha-D-arabinohexopyranoside--Compound 10(IC 83 1373)

It is obtained by the usual method from compound 9.

ANALYSIS C₁₁ H₂₀ ClN₃ O₅ : 309.75--Calculated % C: 42.6, H: 6.5, N:13.6--Found % C: 42.3, H: 6.2, N: 13.6. MP: 60° C. [α]_(D) ²⁰ : +68.8°(c: 0.5% CHCl₃)--[α]₃₆₅ ²⁰ : +113.0° (c: 0.5%, CHCl₃)

NMR Spectrum (DMSO-d₆): 1.19 (d CH₃ J=6 Hz), 1.87 (m CH₂ -2') 3.01 (tH-4'J=J'=9 Hz) 3.25 (s OCH₃ -1) 3.35 (s OCH₃ -4) 3.53 (m H-5) 3.63 (tCH₂ -4 J=J'=6 Hz) 3.61 (m CH₂ -5) 4.18 (m H-3') 4.66 (d H-1' J=3 Hz)8.73 (d NH J=9 Hz).

Mass Spectrum (chemical ionisation): m/e 310 (Cl₃₅, (M+1, Basic peak)and m/e 312 (Cl₃₇, 30%); m/e 278 (m+1-32, 90%) and m/e 280 (27%).

EXAMPLE 14 Methyl 3,6-diazido 4-O-benzyl 2,3,6-tridesoxyα-D-arabinohexopyranoside Compound 11

6 g (0.092 mole) of sodium azotide were added to a solution of 6 g(0.016 mole) of methyl 3-azido 4-O-benzyl 6-bromo 2,3,6-tridesoxyα-D-arabinohexopyranoside in 50 ml of anhydrous dimethylformamide. Thereaction medium is brought to 80° C. for 8 hours. After cooling anddilution with 150 ml of water, it is extracted several times with ethylether. The ether phase, after evaporation to dryness, under vacuum,gives a colorless oil: 5 g (98%).

ANALYSIS C₁₄ H₁₆ N₆ O₄ : 332.32 [α]_(D) ²⁰ : +51° (c: 2.2%, CHCl₃)

IR Spectrum: 3450 cm⁻¹ (OH) 2115 cm⁻¹ (N₃)

EXAMPLE 15 Methyl 3,6-diazido 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 12

5 g (0.015 mole) of compound 11 were placed in solution in 50 ml ofanhydrous methanol, then a molar solution of sodium methylate (50 ml)was added and it was stirred for 3 hours at ambient temperature. Thesolution was neutralized by filtration on Amberlite IK 50 H⁺ resin, thenevaporated to dryness under reduced pressure. A syrupy oil (3.4 g 100%)was obtained which was purified on a silica column H (eluant CH₂ Cl₂).

ANALYSIS C₇ H₁₂ N₆ O₃ [α]_(D) ²⁰ : +128° (c: 1.6%, CHCl₃)

IR Spectrum_(film) : 3450 cm⁻¹ (OH) 2115 cm⁻¹ (N₃)

EXAMPLE 16 Methyl 3,6-diamino 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 13

3 g (0.013 mole) of compound 12 in solution in 100 ml of ethanol with 1%triethylamine are hydrogenated at ordinary pressure for 12 hours in thepresence of palladium on carbon as catalyst. After removal of thecatalyst, the filtrate evaporated to dryness gives a syrupy residue (2.8g). Chromatography on silica (eluant CH₂ Cl₂, ammoniacal MeOH 80:20)enabled isolation of the pure product 13.

ANALYSIS C₇ H₁₆ N₂ O₃ : 176.25 [α]_(D) ²⁰ : +130° (c: 1.13%, MeOH)

IR Spectrum_(film) : 3700 cm⁻¹ (OH), 3360 cm⁻¹ (NH)

EXAMPLE 17 Methyl bis 3,6-[3-(2-chloro ethyl) 3-nitroso ureido]2,3,6-tridesoxy α-D-arabinohexopyranoside.--Compound 14 (IC 83-1374)

It is obtained by the usual method from compound 13.

ANALYSIS C₁₃ H₂₂ Cl₂ N₆ O₇ : 445.267--Calculated % C: 35.1, H: 5.0, N:18.9--Found % C: 35.3, H: 5.1, N: 19.2. MP: 102° C. [α]_(D) ²⁰ : -59.0°(c: 0.5%, CHCl₃)--[α]₃₆₅ ²⁰ : +74.0° (c: 0.5%, CHCl₃)

NMR Spectrum (DMSO-d₆): 1.84 (m CH₂ -2') 3.15 (s OCH₃) 3.17 to 3.77 (mCH₂ -6', H-3', H-4', H-5', 2CH₂ -4) 4.06 (m 2CH₂ -5) 4.66 (d H-1 J=3 Hz)8.41 (t NH J=J'=6 Hz) 8.50 (d NH J=9 Hz).

Mass Spectrum: (M+1): 445 100% Basic peak

    ______________________________________                                        447 Cl 35            449 Cl 37                                                Cl 37                Cl 37                                                    ______________________________________                                    

EXAMPLE 18 Methyl 3-O-acetyl 4,6-O-benzylidene 2-desoxyα-D-arabinohexopyranoside--Compound 15

40 ml of redestilled acetic anhydride are added to 10 g (0.037 mole) ofmethyl 4,6-O-benzylidene 2-desoxy α-D-arabinohexopyranoside in solutionin 50 ml of anhydrous pyridine. After 48 at 50° C., the reaction mixtureis cooled then poured onto cracked ice, extracted 3 times with 100 ml ofdichloramethane. The organic phase dried over sodium sulfate gave afterevaporation, 11.5 g (99%) of a crystalline product that is purified byrecrystallization in the hexane-acetone mixture.

MP: 125°-127° C. [α]_(D) ²⁰ : +74° (c: 1%, chloroform)

IR Spectrum_(Nujol) : 1728, 1240 cm⁻¹ (c=0 ester).

EXAMPLE 19 Methyl 3-O-acetyl 4-O-benzoyl 6-bromo 2,6-didesoxyα-D-arabinohexopyranoside--Compound 16

7.13 g (0.04 mole) of barium carbonate and 3.23 g (0.020 mole) ofN-bromo-succinimide were added to 5.12 g (0.02 mole) of compound 15 insolution in 200 ml of carbon tetrachloride were added. The reaction wasbrought to reflux for 3 hours. After cooling and removal of theinsoluble by filtration, the organic phase was washed with a saturatedsolution of sodium bicarbonate.

After evaporation, 6.4 g (98%) of a single spot oily product in t.l.c.are obtained. [α]_(D) ²⁰ : +70 (c: 1.45%, CHCl₃)

IR Spectrum: 1730 cm⁻¹, 1240 cm⁻¹ (CO ester) 1600, 1585 cm⁻¹ (aromatic)

EXAMPLE 20 Methyl 3-O-acetyl 6-azido 4-O-benzoyl 2,6-didesoxyα-D-arabinohexopyranoside.--Compound 17

To a solution of 8 g (0.020 mole) of compound 16 in 50 ml of anhydrousdimethylformamide were added 8 g (0.12 mole) of sodium azotide. Thereaction mixture was brought to 80° C. for 8 hours.

After cooling and dilution with water, the reaction mixture wasextracted with ether. The solution washed several times with water, wasdried over sodium sulfate. After evaporation under vacuum, the residuewas purified by chromatography on silica (eluant hexane-ethyl acetate,3-1). 6.5 g of pure product were isolated (95%). The product wasrecrystallized from hexane.

ANALYSIS C₁₆ H₁₉ N₃ O₅ : 349.38 MP: 68° C. [α]_(D) ²⁰ : +90 (c: 1%;CHCl₃)

IR: 2100 cm⁻¹ (azide) 1725, 1280, 1050 cm⁻¹ (ester) 1610, 1590 cm⁻¹(aromatic)

EXAMPLE 21 Methyl 6-azido 2,6-didesoxyα-D-arabinohexopyranoside--Compound 18

A methanolic solution of sodium methylate (20 ml) was added to asolution of 4.96 g (0.014 mole) of compound 17 in 50 ml of anhydrousmethanol.

After 12 hours stirring at room temperature, the reaction medium wasneutralized by filtration on Amberlite IR 50 resin (form H⁺). Afterremoval of the solvent, the syrupy residue obtained was chromatographedon a silica column to remove the methyl benzoate. 2.76 g of compound 18were obtained (96%).

ANALYSIS: C₇ H₁₃ N₃ O₄ : 203.2 [α]_(D) ²⁰ : +104° (c: 1%, chloroform).

IR Spectrum: 3400 cm⁻¹ (OH) 2120 cm⁻¹ (azide).

EXAMPLE 22 Methyl 6-amino 2-desoxy α-D-arabinohexopyranoside--Compound19

A solution of 2.40 g (0.011 mole) of compound 18 in 25 ml of anhydrousethanol was shaken under a hydrogen atmosphere in the presence of 10%palladium on carbon (500 mg) for 12 hours. After elimination of thecatalyst, the evaporation of the filtrate led to an oily product 2 g(95%). A sample was converted into the picrate.

ANALYSIS: C₁₃ H₁₈ N₄ O₁₁ : 406.35 MP: 156° C. (ethanol) [α]_(D) ²⁰ :+75° (c: 1.2%, chloroform).

EXAMPLE 23 Methyl 3-[3-(2-chloro ethyl) 3-nitroso ureido] 2,6-didesoxyα-D-arabinohexopyranoside--Compound 20 (IC83 1350)

It is obtained by the usual method from compound 19.

ANALYSIS C₁₀ H₁₈ ClN₃ O₆ : 311.728--Calculated %: C: 38.5, H: 5.8, N:13.5--Found %: C: 38.5, H: 5.5, N: 13.4. MP: 101° C. (dec.) _(D) ²⁰ :+26.2 (c: 0.5%, CHCl₃) ₃₆₅ ²⁰ : +51.0 (c: 0.5%, CHCl₃)

NMR Spectrum (CMSO-d₆): δ 1.45 (t(d) H-2'_(ax) J=J'=12 Hz, J"=4 Hz),1.85 (d(d) H-2'_(eq) J=12 Hz J'=4 Hz), 2.95 (t H-4' J=J'=9 Hz), 3.10(s-OCH₃), 3.30 (m H-3'), 3.51 (m H-5+a CH₂ -6'), 3.58 (t CH₂ -4 J=J'=6Hz), 3.75 (d_(b) CH₂ -6' J_(gem) =12 Hz), 4.06 (m CH₂ -5), 4.15 (dH-1'J=3 Hz), 4.81 (OH), 5.10 (OH), 8.50 (t NH).

Mass Spectrum: (M+1)=312 Cl₃₅ 314 Cl₃₇ (Loss of MeOH -32) 280.

EXAMPLE 24 Methyl N methoxycarbonyl 3-amino 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 21

6 ml of methyl chloroformate were added at 0° C. and with stirring, in10 minutes to 1.5 g (0.0093 mole) of methyl 3-amino 2,3,6-tridesoxyα-D-arabinohexopyranoside in 200 ml of anhydrous methylen chloride. Thereaction mixture was kept 2 hours at ordinary temperature, then 100 mlof 4N soda were added. After one night with stirring, the organic phasewas separated by decantation, the aqueous phase extracted with 100 ml ofmethylene chloride. The organic phase washed with distilled water wasdried over sodium sulfate.

After evaporation under vacuum, the residue obtained was recrystallizedin a methanol-methylene chloride mixture, 1.7 g (80%).

ANALYSIS C₉ H₁₇ NO₅ : 219.24 MP: 180° C. [α]_(D) ²⁰ : +157 (c: 1%,CHCl₃).

EXAMPLE 25 Methyl 3-methylamino 2,3,6-tridesoxyα-D-arabinohexopyranoside--Compound 22

1.7 g (0.0077 mole) of compound 21 in 50 ml of anhydrous ethyl etherwere added drop by drop, so as to maintain a slight reflux (30') to 1 gof lithim and aluminium hydride in 100 ml of anhydrous ethyl ether. Thereflux was continued for 6 hours. After cooling, the excess of hydridewas decomposed by the very slow addition of 1 ml of water, then 1 ml of3N soda, then 3 ml of water. The precipitate was removed by filtration.After evaporation under vacuum and then recrystallization in a mixtureacetone-hexane the organic phase dried over sodium sulfate gave, 1.05 gof crystals (80%); MP: 105° C.

ANALYSIS C₈ H₁₇ NO₅ : 175.23 [α]_(D) ²⁰ : 89 (c: 1%, CHCl₃)

EXAMPLE 26 Methyl 3-[3-(2-chloro ethyl) 1-methyl 3-nitroso ureido]2,3,6-tridesoxy α-D-arabinohexopyranoside--Compound 23 (IC 83.1375)

It was prepared according to the usual method from compound 22.

The examples 5 to 25 above are illustrated by the following reactiondiagrams: ##STR206##

EXAMPLE 27: Methyl 3 [3-(2-chloro ethyl) 3-nitrosoureido]2,3,6-tridesoxy α-L-arabinohexopyranoside Compound 26 -IC 841530

The preparation of methyl 3-amino 2,3,6-tridesoxyα-L-arabinohepoxyranoside (L-acosamine) is first carried out, asmentioned hereafter.

3.1 g (0.04 mole) of anhydrous sodium acetate and 2.1 g (0.024 mole) of0-methyl hydroxylamine chlorhydrate are added to 1.92 g (0.012 mole) ofmethyl 2,6-didesoxy α-L-erythrohexopyranoside 3-ulose in 25 ml of 50%aquous ethanol. The reaction medium is brought to reflux 3 hours, thenethanol is evaporated. After extraction with dichloromethane and dryingover sodium sulfate sodium, a clear oil (2 g) is obtained, the structureof which is confirmed by NMR and corresponds to 0-methyloxime ofL-acosamine.

This oil is solubilized in 20 ml of anhydrous tetrahydrofurane and 30milliequivalents of diborane are added to the solution under nitrogenand at 0° C.

The reaction mixture is brought to reflux two hours, then cooled at 0°C., 5 ml of water, then 5 ml of 20% potash are added carefully. Thereaction medium, brought to reflux 3 hours, then cooled, is extractedwith ethyl acetate. The organic phase gives, after evaporation, aresidue, which, after purification, gives L-acosamine crystals.

M.P. 130°-133° C.

[α]_(D) ²⁰ : -140° C. (c: 0,6%, MeOH).

The preparation of methyl 3-[3(2-chloro ethyl)ureido] 2,3,6-tridesoxyα-L-arabinohexopyranoside (compound 25) is carried out, as mentionedhereafter.

0.3 ml (4.10⁻³ mole) of 2-chloro ethyl isocyanate are added to asolution of 0.48 g (3.10⁻³ mole) of methyl 3-amino 2,3,6-tridesoxyα-L-arabinohexopyranoside in 8 ml of redistillated dimethylformamide.After 2 hours of stirring, the reaction mixture is evaporated to drynessunder vacuum. The crystals are dried, then washed with ether: 781 mg(98%)

NMR Spectrum: Solvent DMSO D₆ : 1,14 (d, CH₃ -6', J=6 Hz); 1,46 (T(d),H-2'_(ox), J=J'=12 Hz, J"=4 Hz); 1,90 (dd,H-2'_(eq), J=12 Hz, J'=4 Hz);2,78 (T,H-4', J=J'=9 Hz); 3,19 (s,OCH₃); 3,28 (m,CH₂ -4); 3,42 (m,H-5');3,53 (m,CH₂ -5); 3,61 (m,H-3'); 4,58 (d,H-1', J=3 Hz); 5,99 (d,NH, J=8Hz); 6,14 (t,NH, J=J'=6 Hz).

The final product which is desired (compound 26), i.e. methyl 3[3-(2-chloroethyl)3-nitroso ureido] 2,3,6-tridesoxy2,3,6α-L-arabinohexopyranoside is obtained from the product which hasbeen previously synthesized, as follows 0.66 g (2.5×10⁻³ mole) of [3(2-chloro ethyl)ureido]2,3,6-tridesoxy α-L-arabinohexopyranoside aredissolved in 5 ml of formic acid. 1.4 (0.02 mole) of sodium nitrite areadded by small portions and under stirring to the solution which is keptat 0° C. After 30 minutes, 5 ml of water are added and the stirring ismaintained for one hour. The reaction mixture is poured on to 100 ml ofethyl acetate then dried over sodium sulfate and evaporated to drynessunder vacuum. After purification, on a silica support column, eluent CH₂Cl₂ : 98, MeOH: 2 colorless crystals are obtained: 210 mg (30%) MP: 100°C.

ANALYSIS C₁₀ H₁₈ ClN₃ O₅ : 295,71.

Calculated %: C: 40,6; H: 6,1; N: 14,2.

Found %: C: 40,9; H: 6,1; N: 13,90.

NMR Spectrum (DMSO,D₆): 1,15 (d,CH₃ -6' J=6 Hz); 1,78 a 1,94 (m, CH₂-2'); 3,04 (t,H-4', J=9 Hz); 3,25 (s,OCH₃); 3,51 (m,H-5'); 3,60 (T,CH₂-4, J=J'=6 Hz); 4,10 (m,CH₂ -5, H-3'); 4,65 (d,H-1',J=3 Hz); 8,48 (d,NH,J=9 Hz).

The reaction diagram hereafter summarizes the steps which have just beendescribed: ##STR207##

Pharmacological study

In order to test the antitumoral activity of the compounds of formula Iin the first place leucemia murin L1210 was used. Among murin leucemias,leucemia L1210 is resistant and selective. A substance having a greatactivity on leucemia L1210 presents a potential of activity in theclinical field of humans (J. M. Venditti, Relevance of transplantableanimal tumor systems to the selection of new agents for clinical trialin pharmacological basis of cancer chemotherapy, the University of Texased Williams and Wilkins Co. publ. 1975, Baltimore U.S.A., p. 245-270).

Besides, the experimental tumor, leucemia L1210 of the mouse is in factcurrently used for the evaluation of all antitumoral compounds atpresent used in human therapy, as described, for example by C. C. Zubrodin Proc. Nat. Acad. Sci. U.S.A., 69, 1972, p. 1 042-047. The tumoralsystem so-constituted experimentally enables very accurate experimentalevaluation of the activity of the compound tested and, consequentlyalso, an objective comparison between the respective activities of thedifferent compounds, for example according to the methods described byR. E. Skipper, F. M. Schapel Jr. and W. S. Wilcox in Cancer Chemother,Rep., 35, 1964, p. 1-111 and 45, 1965, p. 5-28.

This has been confirmed by the results of recent work of Staquet et al.Cancer Treatment Reports, vol. 67, no 9, September, 83.

In practice, the biological effects of the novel nitrosourea derivativesaccording to the present invention have been tested as follows.

METHOD

The test used is that of W. J. Durkin et al. Cancer Research 1979, 39,402-407, modified.

All the nitrosoureas were dissolved in 70% ethanol in the proportion of10 mg/ml.

The test was carried out in two steps.

1--Determination of the 20% cytotoxic index

100 μl of an L 1210 cell suspension (10⁶ cells per ml) in R P M I 1640culture medium supplemented with 10% of fetal calf serum and 40 μg/ml ofgentamycin, containint various doses of the products to be tested (0 to100 μg/ml) were incubated 24 hours at 37° C. At the end of this time,the cell viability is determined by the trypan blue exclusion test. Thecytotoxic index is defined by the formula: ##EQU1##

The amount of ethanol is the same in the cultures containing theproducts to be tested and in the control cultures (this amount has noeffect either on growth nor on cell viability).

For each product the dose which gives a cytotoxic index of 20% isdetermined.

2--Determination of the potential "in vivo" activity

L 1210 cells, under conditions similar to the preceding protocol, werecontacted for 1 hour with a dose of the various products testedcorresponding to a cytotoxic index equal to 20%. After this time, thecells were placed in culture medium not containing nitrosoureas andincubated at 37° C. After 48 hours, the cytotoxic index was determined.

W. J. DURKIN et Al . . . showed that, under these conditions, if thecytotoxic index was equal to or greater than 40%, the product concernedwould be active "in vivo" in the mouse.

RESULTS

The results are collected in the following table I.

                                      TABLE I                                     __________________________________________________________________________               TEST N° 1                                                              DOSE                                                                          CORRESPONDING                                                                 TO A       TEST N°2                                                    20% CYTOTOXIC                                                                            CYTOTOXIC                                                                             POTENTIAL                                       PRODUCT    INDEX      INDEX   "IN VIVO"                                                                            "IN VIVO"                                TESTED     μg/ml   %       ACTION ACTIVITY                                 __________________________________________________________________________    COMP. 2                                                                             IC 1183                                                                            50         77      +      +                                        COMP. 6                                                                             IC 1184                                                                            20 a 25    N.D.           N.D.                                     COMP. 10                                                                            IC 1373                                                                            80         68      +      N.D.                                     COMP. 14                                                                            IC 1374                                                                            5C         71      +      N.D.                                     COMP. 20                                                                            IC 1350                                                                            N.S.       N.S.    -                                               COMP. 23                                                                            IC 1375                                                                            60         69      +      N.D.                                     __________________________________________________________________________     N.D. = undetermined                                                           N.S. = not significant                                                   

3--Determination of the effective "in vivo" activity in the mouse

The experimentation which is reported below used the compound 2 preparedaccording to the above example 6 (Ref. IC 81 1183).

3.1. Protocol

The mice (female, average weight 20 g) used were F₁ /DBA₂ /C₅₇ /B1(Animal selection and breeding center of the Laboratories of the CNRS,Orleans, La Source).

The mice distributed in cages by drawing lots were inoculated on day "0"with 10⁵ leucemia L1210 cells.

The animals were treated with the compound IC 81 1183 intraperitoneallyon days 1, 5 and 9.

The suspensions were prepared just before the injection: product2+neutralized and sterilized olive oil.

The mortality of the animals was observed regularly, the relativeincrease in the survival (T/C×100) was calculated from the averagesurvival of the treated animals (T) and that of the control animals (C).

The doses used are in mg/kg of mouse: 1.25, 2.5, 5, 10, 20, 40, 50, 60,80.

3.2. Results

1.25 mg→T/C=118

2.50 mg→T/C=158

5 mg→T/C=170

10 mg→T/C=220

20 mg→T/C=00

40 mg→T/C=00

50 mg→T/C=100

≧60 mg→Toxicity

3.3. Remarks

(a) T/C=00: for definition: more than 50% of the treated animals werefinally cured; now in the experiments carried, all the animals at dosesof 20 and 40 mg/kg were cured finally.

(b) This curve of efficiency is as good as that obtained with RFCNU andRPCNU described by IMBACH et al (Loc. Cit) and it is very distinctlyhigher than those obtained by CCNU and Me CCNU described by MATHE andKENIS (Loc. cit).

(c) A sudden drop of the T/C is observed after 40 mg/kg. This phenomenonis also observed with nitrosoureas used as comparison products: therapid rise in toxicity cancelled the efficiency of the product.

3.4. The value of T/C≦125% was sought in order to determine a minimumactive dose which is situated between 1.25 mg/kg of body weight and 2.5mg/kg; in fact, at 1.25 mg/kg it is found to be slightly below thesignificant threshold of 125% survival, which is not the case at thedose of 2.5 mg/kg.

4. Determination of the "in vivo" activity of the compounds according tothe invention on the three respective tumors leucemia L 1210 IGR, Lewistumor and melanoma B16.

The compounds tested were the compounds of examples referenced by IC1183, IC 1184, IC 1350, IC 1373 and IC 1374.

4.1. Leucemia L1210

Female DBA₂ mice were used, about 8 weeks old and weighing about 20 gfrom the IFFA-CREDO center (les Oncins, 69210 Arbresles)

At day "0", each mouse received intraperitoneally an inoculum of 1×10tumor cells in a volume of 0.2 ml.

After tumoral graft, the mice were distributed at random into 21 cagesof 5 animals, themselves then distributed, by drawing lots, into 6experimental series. Within these 6 experimental series, there wereconstituted a control series of 6 cages and 5 experimental series of 3cages each, and of which the mice were intended to be treated by thecompounds of the invention.

4.2. Melanoma B16

Female C57 B 1/6 mice were used, about 8 weeks old and weighing about 20g, from the IFFA-CREDO center.

At day "0", each mouse received intraperitoneally an inoculum of 2×10⁶tumor cells in a volume of 0.5 ml.

After tumoral graft, distribution at random was carried out as indicatedabove, to obtain a control series of 8 cages and 5 experimental seriesof 4 cages each, and of which the mice were intended to be treated bythe compounds of the invention.

4.3. Lewis tumor

Procedure was under the conditions which have just been describedpreviously with regard to melanoma B16, by injecting the tumor cells, inthe proportion of 2×10⁶ per mouse, in the volume of 0.2 ml.

4.4. Protocol and treatment

The protocol was identical for the 3 tumors and the 5 compounds of theinvention to be tested.

As regards the experimental series to be treated, each mouse received,at days 1, 5 and 9, intraperitoneally, a dose of 20 mg/kg of thecompound to be tested, in the volume of 0.2 ml of neutralized andsterilized olive oil.

As regards the control series, each mouse received, intraperitoneally,0.2 ml of neutralized and sterilized olive oil.

4.5. Results

Control series (comparison)

The average survival of the comparison mice is expressed in days±2typical deviation of the mean (±26 σ m):

L1210: 8, 9±0.26

3LL Lewis tumor: 12.87±0.79

Melanoma B16: 14.52±0.80.

Treated series (see Table II below)

The results are expressed by the relative increase in survival (T/C×100)calculated from the average survival of the treated animals (T) and thatof the control animals (C)

The mice surviving more than 60 days were considered as cured.

The figures between parentheses indicate the percentage of cured mice.

The sign 00 indicates that 50% at least of the mice were cured.

When the percentage of mice cured is less than 50%, the cured mice areconsidered as dead at 60 days for the calculation of the T/C×100 (inthis case the T/C×100 is hence more or less underestimated).

Among the mice which had lived at least 60 days, and hence considered ascured, certain were killed at the 60th day. No anomaly was observed inmacroscopic examinations of the organs removed.

The other mice were preserved and kept alive to the 200th day; they didnot manifest any apparent disturbance in their behavior.

The animals which died in the course of the experiment were autopsiedand there was no death through toxicity.

                  TABLE II                                                        ______________________________________                                        Compound tested                                                               Tumor   I.C. 1183                                                                              I.C. 1184                                                                              I.C. 1350                                                                            I.C. 1373                                                                            I.C. 1374                             ______________________________________                                        Leucemia                                                                              ∞.sub.(100)                                                                      556.sub.(47)                                                                           362.sub.(20)                                                                         ∞.sub.(100)                                                                    ∞.sub.(67)                      1210                                                                          Tumor   ∞.sub.(75)                                                                       ∞.sub.(90)                                                                       325.sub.(40)                                                                         316.sub.(30)                                                                         ∞.sub.(75)                      LEWIS 3LL                                                                     MELA-   234.sub.(5)                                                                            235.sub.(15)                                                                           225.sub.(5)                                                                          165.sub.(0)                                                                          246.sub.(10)                          NOMA B16                                                                      ______________________________________                                    

5 Determination of the "in vivo" activity of the compounds according tothe invention on the tumors L1210 U.S.A. and L1210 IGR.

The tumor L1210 U.S.A. is more resistant than tumor L1210 IGR.

The compounds tested were compounds IC 1183, IC 1184, IC 1350, IC 1373and IC 1374.

The experiments were identical to those described in preceding paragraph4 but lower doses were used.

There were inoculated into the mice, 10⁵ tumor cells of L1210 U.S.A.intraperitoneally in a volume of 0.2 ml.

At days 1, 5 and 9, the compounds of the invention under test wereinjected intraperitoneally, at a lower dose than that used in theexperiment described at 4, that is to say in the proportion of 5 mg/kg.

The same protocol was carried out with the tumor L1210 IGR, byinoculating 10⁵ tumor cells of L1210 IGR intraperitoneally in the volumeof 0.2 ml then, at days 1, 5 and 9, there were injectedintraperitoneally 5 mg/kg of each of the compounds to be tested.

The results are shown in Tables III and IV below.

                                      TABLE III                                   __________________________________________________________________________    L 1210 USA Tumor                                                                       (Comparison)                                                                  Controles                                                                            IC 1350                                                                             IC 1373                                                                             IC 1374                                                                             IC 1183                                                                             IC 1184                               __________________________________________________________________________    Average survival                                                                       8,6 ± 0,22                                                                        10,5 ± 0,74                                                                      10,6 ± 0,44                                                                      12,9 ± 0,55                                                                      13,5 ± 0,91                                                                      18 ± 1,19                          ±2 σm                                                                Variance 0,013  0,139 0,049 0,077 0,205 0,355                                 (S.sub.m.sup.2 )                                                              T/C × 100 126   123   150   157   209                                   Median survival                                                                        8,7    10,25 10,6  13,1  14,0  17,8                                  T/C × 100 118   122   151   161   205                                   __________________________________________________________________________

                                      TABLE IV                                    __________________________________________________________________________    L 1210 IGR TUMOR                                                                       (Comparison)                                                                  Controles                                                                            I.C. 1350                                                                           I.C. 1373                                                                           I.C. 1374                                                                           I.C. 1183                                                                           I.C. 1184                             __________________________________________________________________________    Average survival                                                                       9,05 ± 0,18                                                                       14,3 ± 1,23                                                                      12,9 ± 0,55                                                                      17,1 ± 2,10                                                                      18,7 ± 1,30                              ±2 σm                                                                Variance 0,008  0,379 0,077 1,10  0,423                                       (S.sub.m.sup.2 )                                                              T/C × 100 158   142   189   207   378                                   Median survival                                                                        9,05   14,25 13,1  16,75 19    24                                    T/C × 100 157   145   185   210   265                                   __________________________________________________________________________

6. Study of the toxicty of the compounds according to the invention

Histological examinations were carried out on the organs of DBA₂ miceinoculated by the tumor L1210 IGR and treated for 60 days with productsIC 1183, IC 1184, IC 1350, IC 1373 and IC 1374, as well as on organs ofthe comparison DBA₂ mice, that is to say inoculated with the tumor L1210IGR.

The organs subjected to these histological examinations were the liver,the kidneys, the spleen, the adrenal glands and the lungs.

Examination showed that the comparison animals inoculated by L1210 IGRpresented a cellular disorganisation of the hepatocytes.

On the contrary, the hepatic structure of mice inoculated by L1210 IGR,then treated with the products according to the invention, did not showas great a disturbance as that observed with the comparison miceinoculated with L1210 IGR, and the hepatic nuclear hypertrophy does notconsequently seem to be connected with the toxicity itself of theproducts of the invention.

A hematological study has been carried out on some animals treated withthe products according to the invention; it is the case particularly ofDBA₂ mice inoculated with L1210 leucemia and C57/B16 mice which havebeen inoculated with Lewis tumor. The study comprised, from a bloodsampling by cardiac puncture, collected on heparin, a blood count(erythrocytes and leukocytes), hematocrit, platelet count, differentialblood count.

From marrow smears of femoral origin and from a spleen print, a shortstudy of hematopoietic centers has been undertaken.

A histological study has also been carried out on liver, spleen,kidneys, adrenal glands, lungs, which were sampled when the animals werekilled.

No important disturbances have been observed, either in the blood countor in the differential blood count; there is no bone marrowaplasia andthe marrows which have been observed are rich in cells of all kinds. Thespleens seem to be substantially normal.

7. General conclusion

The animal experiments carried out with the product according to theinvention give interesting results when the model selected in melanomaB16, and excellent results when the models selected are the Lewis tumorand the L1210 IGR tumor as well as the L1210 U.S.A. tumor, moreresistant than the L1210 IGR.

The compounds according to the invention are hence particularly suitablefor the treatment of various human cancers, especially those which aresensitive to chimiotherapy. The compounds of the invention areparticularly suitable for the treatment of various forms of cancermeeting this condition and which are identified in the publicationsalready mentioned. The compounds of the invention are also suitable forthe treatment of primary and secondary cerebral tumors,broncho-pulmonary tumors, tumors of the ORL sphere, digestive tumors(gastric, pancreatic, colic and rectal), tumors of the breast, of thegenital organs in the woman, bone tumors (osteosarcomas,reticulosarcomas), melanomas, hemato-sarcomas (Hodgkinian and nonHodgkinian lymphomas), and multiple myelomas.

The invention relates also to pharmaceutical compositions comprising theabove said novel compounds in association with a pharmaceutical vehiclesuitable for the selected mode of administration.

The invention relates particularly to sterile or sterilizable solutions,injectable or suitable for use for the preparation, particularlyextemporaneously, of injectable solutions suitable for administration byintravenous injections or perfusions. They relate, in particular, tophysiologically acceptable hydroalcoholic solutions.

The products according to the invention, may be for instance presentedin the form of freeze-dried powder, which, for administration, isprepared extemporaneously by solubilisation by means of a sterilealcoholic solvent. The solution so-obtained is then diluted withapyrogenic sterile water, then before being administered by intravenousperfusion, the solution is again rediluted in 9% isotonic salt serum or5% isotonic glucose serum.

The doses administered daily must be sufficient so that an action can bemanifested at least in a relatively large proportion of patientsafflicted with one or other of the various forms of cancer which are orwill be accessible to chimiotherapy, however without nonethelessexceeding those for which the compounds become too toxic.

More particularly, the doses to be administered are determined accordingto models conventionally used in this field which are, for example,described in the two following articles:

Cancer Research, 37, 1 934-1 937, June 1977, P. S. Schein;

Cancer Chimiotherapy Reports, vol. 50, no 4, May 1966, E. J. Freireich.

The model for determining the suitable doses for a given compound,consist of determining the dose which is tolerated by the animal andwhich corresponds to about 1/10 of the lethal dose (LD10) expressed inmg/m2 of body surface. The doses which can be used in man correspond to1/3 to 1/10 of the LD10 dose mentioned above (cf. Cancer Research, 37,1935, column 1, June 1977).

By way of example, the daily doses administered by the general route,particularly by perfusion, and expressed in mg/kg can vary from about 1to about 50 mg/kg, for example, about 3 mg/kg.

The invention also relates to other forms of administration, especially,for the oral route (solid or liquid compositions) or for the rectalroute (glycerin compositions suitable for the latter route).

These dosage ranges are obviously only by way of indication. It isnaturally understood that in this type of therapy, the dosesadministered must in each case be evaluated by the clinician taking intoaccount the state of the patient and of his personal reactivity withrespect to the medicaments.

An example of pharmaceutical composition of the products according tothe invention comprises 100 mg of at least one of the products of theinvention, presented in the form of a sterile freeze-dried powder,associated with an ampoula of physiologically acceptable solvent,particularly of alcohol, such as ethanol, at the dosage of about 5 mlper ampoula.

Because of their particularly important activity, the compounds of theinvention are also useful as reference products in pharmacologicalstudies, particularly in order to carry out antitumor comparisons of theproducts which are studied with respect to a reference product.

Example 28

Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4-dideoxy-β-D-xylo-hexopyranoside (CI. 1675).

(1) Preparation of methyl 3-azido 6-benzoyl3-deoxy-β-D-gluco-hexopyranoside

29 g of 3-azido 3-deoxy D-gluco-hexopyranoside are dissolved in 600 mlof 1N hydrogen chloride in methanol, the solution is heated under refluxfor 2 hours and then evaporated to dryness in a vacuum.

The product is taken up in dichloromethane and washed with water untilchloride ions have been completely removed. The organic phase is driedover sodium sulfate then evaporated to dryness in a vacuum to give aresidue of 31 g of glycosylated product which is used as such in thenext step.

The crude product is dissolved in 250 ml of anhydrous toluene. 90 ml ofbis tributyl tin oxide are then added and the reaction mixture is thenrefluxed for 3 hours. The reaction mixture is cooled to -15° C. and asolution of 40 ml of benzoyl chloride in 200 ml of methylene chloride isadded dropwise. After being stirred for 24 hours, the reaction mixtureis evaporated to dryness in a vacuum, then chromatographed on silica(eluant: CH₂ Cl₂ :CH₃ OH,98:2).

This leads to the isolation, in the form of resinous substances, of:

7.5 g of methyl 3-azido 6-benzoyl 3-deoxy-β-D-gluco-hexopyranoside.[α]_(D) =-3.5° (c,0.83 CHCl₃)

7.0 g of methyl 3-azido 6-benzoyl 3-deoxy-α-D-gluco-hexopyranoside.[α]_(D) =+102.0° (c,1.14 CH₂ Cl₂).

(2) Preparation of methyl 3-azido 6-benzoyl 4-chloro3,4-dideoxy-β-D-galacto-hexopyranoside

To 4.8 g (0.015 mole) of methyl 3-azido 6-benzoyl3-deoxy-β-D-gluco-hexopyranoside in 250 ml of anydrous pyridine 12.5 ml(0.15 mole) of sulfuryl chloride are added dropwise at 0° C. Thesolution is stirred for 18 hours at 0° C., then allowed to warm to roomtemperature. The reaction mixture is then poured onto 500 g of ice andextracted with dichloromethane.

The organic phase is washed with 1N sulfuric acid, then with water,dried over sodium sulfate, filtered and evaporated. The oily residue ispurified by chromatography on silica using the eluant hexane-ethylacetate 4:1 to give 4.05 g (80%) of white crystals.

C₁₄ H₁₆ ClN₃ O₅ : 341.7

mp: 84°-86° (hexane-ethyl acetate)

[α]_(D) : +2.3° (c,0.88 CHCl₃)

(3) Preparation of methyl 3-amino 6-benzoyl3,4-dideoxy-β-D-xylo-hexopyranoside

To 4.05 g (0.012 mole) of the preceding compound in 200 ml of anhydroustoluene is added azo-bis-isobutyronitrile (700 mg, 4.27 mmoles),followed, under nitrogen and dropwise, by tributyl tin hydride (12.6 ml,47 mmoles). The reaction mixture is refluxed for 2 hours and thenevaporated under reduced pressure.

After chromotography on silica using the eluantdichloromethane-ammoniacal methanol 19:1, 3 g (90%) white crystals areisolated.

C₁₄ H₁₉ NO₅ : 281.3

mp: 112°-117°

[α]_(D) : -13.6° (c,0.3% CH₃ OH)

(4) Preparation of methyl 3-amino 3,4-dideoxy-β-D-xylo-hexopyranoside

To 3 g (0.0107 mole) of the preceding compound in methanol (45 ml) isadded a molar solution of sodium methylate (5 ml). After being stirredfor 2 hours at room temperature the reaction mixture is evaporated underreduced pressure. The residue is purified by chromatography on silicausing the eluant dichloromethane-ammoniacal methanol 17:3.

1.5 g of crystalline product are isolated.

C₇ H₁₅ NO₄ : 177.2

mp: 155°-158° (acetonitrile)

[α]_(D) : -43.5° (c,0.74 CH₃ OH)

(5) Preparation of methyl 3-[3-(2-chloro ethyl) 3-nitroso ureido]3,4-dideoxy-β-D-xylo-hexopyranoside

5×10⁻³ mole of methyl 3-amino 3,4-dideoxy-β-D-xylo-hexopyranoside aredissolved in 2 ml of anhydrous DMF and 5×10⁻³ mole of 2-chloroethylisocyanate are added dropwise at 0° C. with stirring. After beingstirred for 5 hours, the reaction mixture is evaporated to dryness in avacuum. A residue is dissolved in 8 ml of formic acid. 0.036 mole ofsodium nitrite are added in small portions and with stirring to thesolution maintained at 0° C. After 30 minutes 10 ml of water are addedand stirring is continued for 1 hour. The reaction mixture is pouredinto 100 ml of pure ethyl acetate, dried over sodium sulfate andevaporated to dryness. The residue is purified by crystallization fromisopropyl ether.

Analysis: C₁₀ H₁₈ ClN₃ O₆ : 311.7 Yield: 49%

mp: 111°-112°

[α]_(D) : +5.7° (c,1.4% CH₃ OH).

Example 29

Preparation of methyl 3-[3-(2-chloro ethyl)-3 nitroso ureido] 6-benzoyl3,4-dideoxy-β-D-xylo-hexopyranoside (CI. 1674).

Methyl 3-amino 6-benzoyl 3,4-dideoxy-β-D-xylo-hexopyranoside is preparedas described in Example 1, step 3, then the amine obtained is treated asdescribed in example 1, step 5.

Analysis: C₁₇ H₂₂ ClN₃ O₇ : 415.83 Yield: 60%

mp: 108°

[α]_(D) : +7.3 (c,0.28 CH₃ OH)

Example 30

Preparation of methyl 4-chloro 3-[3-(2-chloroethyl) 3-nitrosoureido]3,4-dideoxy-β-D-galacto-hexopyranoside (CI: 1803).

(1) Preparation of methyl 3-amino 6-benzoyl 4-chloro3,4-dideoxy-β-D-galacto-hexopyranoside.

To 3 g of methyl 3-azido 6-benzoyl 4-chloro 3,4dideoxy-β-D-galacto-hexopyranoside in 100 ml of absolute ethanol and 2ml of triethylamine 500 mg of 10% palladium on charcoal are added undernitrogen and the reaction mixture is stirred in an atmosphere ofhydrogen for 24 hours.

After the catalyst has been removed the filtrate is evaporated; theresidue is purified by chromatography on silica.

The product is recrystallized from ethyl ether.

Analysis: C₁₄ H₁₈ ClNO₅ : 315.5 Yield: 82%

mp: 160°-162°

[α]_(D) =-11.0° (c,0.85 CHCl₃).

(2) Preparation of methyl 3-amino 4-chloro3,4-dideoxy-β-D-galacto-hexopyranoside

The debenzoylation is carried out according to the standard procedureusing sodium methylate.

The product obtained has the following properties:

Analysis: C₇ H₁₄ ClNO₄ : 211.5

mp: 158°-162°

[α]_(D) : -4.0° (c,0.84 CHCl₃)

(3) Preparation of methyl 4-chloro 3-[3-(2-chloro ethyl) 3-nitrosoureido] 3,4-dideoxy-β-D-galacto-hexopyranoside.

The preparation was carried out as was described in Example 1, step 5,starting from the compound obtained in step 2 above.

Analysis: C₁₀ H₁₇ Cl₂ N₃ O₆ : 346.17

mp: 145°-148°

[α]_(D) : +30° (c,0.65 MeOH).

Example 31

Preparation of methyl 3-[3-(2-chloro ethyl) 3-nitroso-ureido]3,4-dideoxy-α-D-xylo-hexopyranoside (CI. 1677)

(1) Preparation of methyl 3-azido 6-benzoyl 3-α-D-gluco-hexopyranoside

29 g of 3-azido 3-deoxy D gluco-hexopyranoside are dissolved in 600 mlof 1N methanolic hydrogen chloride and the solution is heated at refluxfor 2 hours, then evaporated to dryness in a vacuum.

The product is taken up in dichloromethane and the solution is washedwith water until chloride ion can no longer be detected. The organicphase is dried over sodium sulfate and evaporated to dryness in a vacuumto give a residue of 31 g of glycosylated product used as such in thenext step.

The crude product is dissolved in 250 ml of anhydrous toluene. 90 ml ofbis tributyl tin oxide are then added and the mixture is refluxed for 3hours. The reaction is cooled to -15° C. and a solution of 40 ml ofbenzoyl chloride in 200 ml of dichloromethane are added dropwise. Afterbeing stirred for 24 hours, the reaction mixture is evaporated todryness in a vacuum and the residue obtained is chromatographed onsilica (eluant CH₂ Cl₂ :CH₃ OH,98:2).

This results in the isolation, in the form of resinous substances, of:

7.5 g of methyl 3-azido 6-benzoyl 3-deoxy-β-D-glucohexopyranoside

7.0 g of methyl 3-azido 6-benzoyl 3-deoxy-α-D-glucohexopyranoside.

(2) Preparation of methyl 3-azido 6-benzoyl 4-chloro3,4-dideoxy-α-D-galacto-hexopyranoside.

To 4.8 g (0.015 mole) of methyl 3-azido 6-benzoyl3-deoxy-α-D-gluco-hexopyranoside in 250 ml of anhydrous pyridine areadded dropwise at 0° C. 12.5 ml (0.15 mole) of sulfuryl chloride. Thesolution is stirred for 18 hours at 0° C., then allowed to warm to roomtemperature. The reaction mixture is then poured onto 500 g of ice andextracted with dichloromethane.

The organic phase is washed with 1N sulfuric acid, then with water,dried over sodium sulfate, filtered and evaporated. The oily residue ispurified by chromatography on silica using the eluant hexane-ethylacetate 4:1 to give 4.05 g (80%) of white crystals.

C₁₄ H₁₆ ClN₃ O₅ : 341.7

mp: 104°-106° (hexane-ethyl acetate)

[α]_(D) : +140.5° (c,1.3 CHCl₃).

(3) Preparation of methyl 3-amino 6-benzoyl3,4-dideoxy-α-D-xylo-hexopyranoside

To 4.05 g (0.012 mole) of the preceding compound in 200 ml of anhydroustoluene is added azo-bis-isobutyronitrile (700 mg, 4.27 mmoles)followed, dropwise and under nitrogen, by tributyl tin hydride (12.6 ml,47 mmoles). The reaction mixture is heated at reflux for 2 hours andthen evaporated under reduced pressure.

After chromatography on silica using the eluantdichloromethane-ammoniacal methanol 19:1, 3 g (90%) of white crystalsare isolated.

(4) Preparation of methyl 3-amino 3,4-dideoxy-α-D-xylo-hexopyranoside

To 3 g (0.0107 mole) of the preceding compound in methanol (45 ml) isadded a molar solution of sodium methylate (5 ml). After being stirredfor 2 hours at room temperature the reaction mixture is evaporated underreduced pressure. The residue is purified by chromatography on silicausing the eluant dichloromethane-ammoniacal methanol 17:3.

1.5 g of crystalline product are isolated.

mp=130°-134° C.

[α]_(D) : +163° (c,0.9 CHCl₃)

(5) Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4-dideoxy-α-D-xylo-hexopyranoside

5×10⁻³ mole of methyl 3-amino 3,4-dideoxy-α-D-xylo-hexopyranoside aredissolved in 2 ml of anhydrous DMF and 5×10⁻³ mole of 2-chloroethylisocyanate are added dropwise at 0° C. with stirring. After beingstirred for 5 hours, the reaction mixture is evaporated to dryness in avacuum. A residue is dissolved in 8 ml of formic acid. 0.036 mole ofsodium nitrite are added in small portions and with stirring to thesolution maintained at 0° C. After 30 minutes 10 ml of water are addedand stirring is continued for 1 hour. The reaction mixture is pouredinto 100 ml of pure ethyl acetate, dried over sodium sulfate andevaporated to dryness. The residue is purified by crystallization fromisopropyl ether.

Analysis: C₁₀ H₁₈ ClN₃ O₆ : 311.7

mp: 109°-110°

[α]_(D) : +120.8° (c,0.48% MeOH).

Example 32

Preparation of methyl 4-chloro 3-[3-(2-chloro-ethyl) 3-nitroso ureido]3,4-dideoxy-α-D-galacto-hexopyranoside (CI. 1676).

(1) Preparation of methyl 3-amino 6-benzoyl 4-chloro3,4-dideoxy-α-D-galacto-hexopyranoside

To 3 g of methyl 3-azido 6-benzoyl 4-chloro3,4-dideoxy-α-D-galacto-hexopyranoside in 100 ml of absolute ethanol and2 ml of triethylamine 500 mg of 10% palladium on charcoal are addedunder nitrogen and the reaction mixture is stirred in an atmosphere ofhydrogen for 24 hours.

After the catalyst has been removed the filtrate is evaporated; theresidue is purified by chromatography on silica.

The product is recrystallized from ethyl ether.

Analysis: C₁₄ H₁₈ ClNO₅ : 315.5

mp: 161°-164°

[α]_(D) : +118.5° (c,1.25 CHCl₃).

(2) Preparation of methyl 3-amino 4-chloro3,4-dideoxy-α-D-galacto-hexopyranoside

The debenzoylation is carried out according to the standard procedureusing sodium methylate.

The product obtained has the following properties:

Analysis: C₇ H₁₄ ClNO₄ : 211.5

mp: 135°-138°

[α]_(D) : +184° (c,0.92 MeOH).

(3) Preparation of methyl 4-chloro 3-[3-(2-chloro-ethyl) 3-nitrosoureido] 3,4-dideoxy-α-D-galacto-hexopyranoside

The preparation was carried out as described in Example 1, step 5,starting from the compound obtained in step 2 above.

Analysis: C₁₀ H₁₇ Cl₂ N₃ O₆ : 346.17

mp: 129°-130°

[α]_(D) : +139.3° (c,0.5 CH₃ OH).

PHARMACOLOGICAL STUDY:

In order to test the antitumor activity of the compounds of formulaIA,--use has primarily been made of murine leukemia L1210. Use has alsobeen made of the tumor melanoma B16. In particular, the products of theinvention have been tested against the melanoma B16 by administeringthem by the I.P. and by the I.V. routes.

The compounds of formula IA--have also been tested against the coloncarcinoma C38 by administering them by the I.P. and I.V. routes.

In practice, the biological effects of the novel nitrosourea derivativescorresponding to the present invention were tested as follows:

PROTOCOL FOR TREATMENT WITH THE PRODUCTS OF THE INVENTION

I. MATERIALS AND METHODS

A. Tumors used

Two murine tumors were used for the "in vivo" studies: the leukemiaL1210 and the melanoma B16.

(1) LEUKEMIA L1210 Animals

The experiments were also performed on female mice which were specificpathogen-free (S.P.F.) of

either the line DBA/2JIco

or line B6 D2 F1/JIco

(first generation hybrids between the lines C57BL/6 and DBA/2). The lineused is specified for each series of experiments.

Tumoral graft

On the day of the graft (by convention day 0=DO) an inoculum of 1×10⁵tumor cells in a volume of 0.2 ml is administered to each mouse by theintraperitoneal route (I.P.).

This inoculum is prepared by diluting ascites fluid taken from theperitoneum of a donor female mouse in NCTC 109 medium (EurobioLaboratories Paris, France), counting of the cells in a MALASSEZ cellunder a microscope and adjustment of the concentration to 5×10⁵ cellsper ml.

Distribution in experimental groups

After the tumor graft has been made, the mice are distributed at randomin cages of 5 animals. Subsequently, these cages containing 5 mice arethemselves distributed at random into a control group (control) andgroups treated with the products of the invention.

(2) MELANOMA B16 Animals

The experiments were always carried out on SPF female mice B6 D2 F1/JIco(first generation hybrids between the lines C5BL/6 and DBA/2).

Tumoral graft

On the day of the graft (Day 0) an inoculum of 2×10⁶ tumor cells in avolume of 0.5 ml is administered to each mouse by the intraperitonealroute (I.P.). This inoculum is prepared from a sub-cutaneous tumorexcised from a donor female mouse. After excision the tumor isfragmented by means of a pair of scissors in the NCTC 109 medium. Afterfiltration through sterile gauze in order to remove large cellularfragments, the homogeneous cell suspension obtained is counted by meansof a MALASSEZ cell and diluted to the desired concentration (4×10⁶ tumorcells per ml) by dilution with the NCTC 109 medium.

Distribution in experimental groups

This was performed in the manner described previously for the leukemiaL1210.

B. Protocol for treatment

The doses of the products of the invention used in the differentexperiments are expressed in milligrams per kilogram of body weight.

The products to be injected were dissolved in isotonic sodium chloridesolution.

Two protocols for treatment were used in the different experiments:

either a single injection on D1, by the intraperitoneal (I.P.) orintravenous (I.V.) route,

or 3 I.P. injections on D1, D5, D9.

In each experiment the animals of the control group received 1 or moreinjections, depending on the experimental protocol used, by the sameroute (I.P. or I.V.) of the same volume (0.2 ml/20 g) of the vehicle notcontaining the active principle (isotonic sodium chloride solution).

II. EXPRESSION OF THE RESULTS

For each experiment a table specifies:

the number and percentage of the total of the mice surviving to D60,

the T/C×100 for treated groups.

T representing the mean survival time of the mice in the treated group

C representing the mean survival time of the mice in the control group(control).

III. COMMENT

The strains of mice used, the experimental protocols and the mode ofexpression of the results are in accordance with directive 271 F of the"N.C.I. Division of Cancer Treatment" (November 1983).

IV. PHARMACOLOGICAL RESULTS

(1) Toxicological study:

The results relating to toxicology are brought together in Table IAbelow:

                  TABLE IA                                                        ______________________________________                                                LD.sub.0    LD.sub.50   LD.sub.90                                     CI      mg/kg       mg/kg       mg/kg                                         ______________________________________                                        1674    ≧40  --          --                                                                approximately                                             1675    ≧20  40          --                                            1676    ≧20  --          --                                            1677    --          --          ≦20                                    ______________________________________                                    

(2) Activity of the products CI. 1674, 1675, 1677 by the I.P. route onthe leukemia L1210 U.S.A. strain and the melanoma B16. Compound of theinvention CI. 1674:

L1210 U.S.A. strain - B₆ D₂ S₁ - I.P. treatment on D1, D5, D9 3×1.25mg/kg, 3×5 mg/kg

    ______________________________________                                                  T/C  Number of survivors at D60                                     ______________________________________                                        3 × 1.25                                                                            111    0/10                                                       3 × 5 130    0/10                                                       ______________________________________                                    

Compound of the invention CI. 1675:

L1210 U.S.A. strain - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×1.25mg/kg, 3×5 mg/kg

    ______________________________________                                                 T/C   Number of survivors at D60                                     ______________________________________                                        3 × 1.25                                                                           164     0/10                                                       3 × 5                                                                              >600    5/10                                                       ______________________________________                                    

Melanoma B16 - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors at D60                                          ______________________________________                                        258       8/10                                                                ______________________________________                                    

Compound of the invention CI. 1676:

L1210 U.S.A. strain - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×1.25mg/kg, 3×5 mg/kg

    ______________________________________                                                  T/C  Number of survivors at D60                                     ______________________________________                                        3 × 1.25                                                                            136    0/10                                                       3 × 5 188    0/10                                                       ______________________________________                                    

Melanoma B16 - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors at D60                                          ______________________________________                                        202       3/10                                                                ______________________________________                                    

Compound of the invention CI. 1677:

L1210 U.S.A. strain - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×1.25mg/kg, 3×5 mg/kg

    ______________________________________                                                 T/C   Number of survivors at D60                                     ______________________________________                                        3 × 1.25                                                                           157     0/10                                                       3 × 5                                                                              >600    6/10                                                       ______________________________________                                    

Melanoma B16 - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors at D60                                          ______________________________________                                        257       7/10                                                                ______________________________________                                    

(3) Activity of the substances of the invention toward melanoma B16after administration by the intravenous route:

The activity of the substances of the invention was also tested byadministering the said substances of the invention by the I.V. route.0.5 ml of an homogenate of the tumor (1 g/10 ml) is administered toB6C3F1 female mice by the subcutaneous (S.C. route) on day 0. Theproducts of the invention and the substances used for comparison areadministered on days 3, 7 and 11 in isotonic solution.

In Table IIA below the results are presented relating to the activity ofthe substance CI. 1675 administered by the I.V. route on the growth ofthe melanoma B16 implanted subcutaneously, in comparison with carmustineBCNU marketed under the name of BiCNU®.

In Table IIA below and in the subsequent tables, T represents the meanweight (or median) of the tumors in the treated mice, C represents themean weight (or median) of the tumors in the control, untreated mice andthe ratio T/C% corresponds to T/C×100.

                  TABLE IIA                                                       ______________________________________                                               Dose     Median weight                                                                             Median weight                                            mg/kg/   of the tumor                                                                              of the tumor                                                                            T/C                                     Product                                                                              inj.     on day 3 (mg)                                                                             on day 30 (mg)                                                                          on day 30                               ______________________________________                                        IC1675 20       36,0        688       8,8                                            15       23,0        1 044     13,4                                           10       32,0        2 025     25,9                                    BCNU   20       40          2 581     33,0                                    Controls                                                                              0       40          7 812     100                                     ______________________________________                                    

In this case it should be noted that the smaller the ratio the better isthe activity, in the ideal case, the weight of the tumor in the treatedmouse is 0 mg and hence the ratio T/C% is equal to 0.

The results obtained show that the substances of the invention areactive when they are administered by the I.V. route, a finding whichmakes them quite suitable for use in human therapy.

(4) Activity of the substance of the invention CI. 1675 administered bythe I.P. route to mice bearing the colon carcinoma C38, implantedsubcutaneously.

A tumor fragment is implanted on day 0 in female B₆ D₂ F₁ mice (19-22g). The treatment with all of the products is administered by the I.P.route on days 2 and 9 (which is a delayed treatment). The weight of thetumors is calculated on day 20.

The results are presented in Table IIIA below.

                  TABLE IIIA                                                      ______________________________________                                                         Weight of                                                                     the tumor on    Number of mice                                      Dose      day 20     T/C  without tumor/total                          Product                                                                              mg/kg/inj.                                                                              (mg)       %    + remarks                                    ______________________________________                                        IC 1675                                                                              30        0          0    7/7 and 5 dead mice                                 20        0          0    7/9                                                 10        272        67   1/10                                         TCNU   30        --         --   10 prematurely dead                                 15        0          0    7/10                                                7.5       266        65   2/10                                         Controls                                                                             0         405        100  0/32                                         ______________________________________                                    

Table IIIA above illustrates the considerable efficacy of the substanceof the invention CI.1675 against the colon carcinoma C38 when it isadministered by the I.P. route by delayed treatment. This is predictiveof activity in man for treatment against various tumors.

(5) Antitumor activity of the substance CI.1675 against the coloncarcinoma C38: comparison of the intravenous and intraperitoneal routes.

With a view to verifying the antitumor activity of the product of theinvention CI.1671 when it is administered by the I.V. route, anotherexperiment was performed in which this product was administered by theI.P. route and the I.V. route for the purposes of comparison.

This experiment is carried out under particularly difficult conditions,the first day of the treatment corresponding to the eighth day after theimplantation of the carcinoma C38, whereas normally the treatment startson the day of implantation of the tumor (D0) or 1 day after (D1) theimplantation of the tumor.

The median weight of the tumors on the first day of treatment (day 8) isabout 12 mg (tumors varying from 8 to 40 mg). The animals are treated ondays 8, 12 and 16 depending on the different routes indicated.

The results relating to the action of the substance of the invention CI.1675 in mice bearing the adenocarcinoma 38 of the colon are broughttogether in Table IVA below.

                                      TABLE IVA                                   __________________________________________________________________________                 I.P. Route        I.V. Route                                                  Median wt.  Mice  Median wt.  Mice                                            of the      without                                                                             of the      without                                   Dose  tumor T/C % tumor/total                                                                         tumor T/C % tumor/total                        Product                                                                              mg/kg/inj.                                                                          D20                                                                              D27                                                                              D20                                                                              D27                                                                              D20                                                                              D27                                                                              D20                                                                              D27                                                                              D20                                                                              D27                                                                              D20                                                                              D27                             __________________________________________________________________________    IC 1675                                                                              20    13 172                                                                              2  13 1/8                                                                              2/8                                                                              0  0  0  0  6/8                                                                              6/6                                    15    88 726                                                                              15 56 0  0  0  0  0  0  7/8                                                                              7/7                                    10    324                                                                              1099                                                                             57 84 0  0  23 352                                                                              4  27 0/8                                                                              0/8                             BCNU   20    196                                                                              274                                                                              34 21 0/8                                                                              0/6                                               CONTROLS                                                                             0     586                                                                              1295                                                                             100   0/24  586                                                                              1295                                                                             100   0/24                               __________________________________________________________________________

It can be seen that not only is the activity of the compounds of formulaIA--towards the carcinoma C38 remarkable but that it is even higherafter administration by the I.V. route than after I.P. administration,since at doses not producing toxicity (15 mg/kg/injection), a totalregression of the tumor can also be observed. In fact, on the 20th--daya tumor, which can be measured at the beginning of the treatment, can nolonger be observed, and on the 27th day there was no sign of recurrence.This is demonstrated by the number of mice without a tumor on the 27thday.

GENERAL CONCLUSION

The experiments in animals performed with the compounds of formula IAgive very interesting results in the models used : tumor L1210 U.S.A.and melanoma B16, and excellent results in the model of the coloncarcinoma C38.

All of the results obtained with these models show that the compounds ofthe invention have a wider spectrum of activity than the other antitumorsubstances known at present.

Furthermore, the compounds of formula IA--offer the remarkable advantageof bringing about the regression of animal tumors very difficult totreat with the known antitumor agents presently available.

Compounds of formula IA are hence--therapeutically active substancesand, from this point of view, they represent another aspect of thepresent invention.

The compounds according to the invention are particularly suited to thetreatment of various human cancers, particularly those which respond tochemotherapy. They are particularly suited to the treatment ofbroncho-pulmonary tumors, tumors of the ENT sphere, digestive tumors(gastric, pancreatic, colic and rectal), breast tumors, genital tumors,bone tumors (osteosarcomas, reticulo-sarcomas), melanomas,hepato-sarcomas (Hodgkin and non-Hodgkin lymphomas), multiple myelomas.

The invention also relates to the pharmaceutical composition containingthe new compounds mentioned above in combination with a pharmaceuticalvehicle appropriate to the chosen mode of administration. The inventionrelates particularly to injectible sterile solutions suitable foradministration by injections or intravenous perfusions. The inventionrelates in particular to physiologically acceptable aqueous-alcoholicsolutions.

The products according to the invention may, for example, be madeavailable in the form of a lyophilized powder, the administration ofwhich requires that it be made up into a solution immediately before usewith the aid of a sterile alcoholic solvent. The solution thus obtainedis then diluted with pyrogen-free sterile water and then, before beingadministered by intravenous perfusion, the solution is diluted againwith a solution of glucose or isotonic sodium chloride.

The doses admininstered must be sufficiently high so as to provide anactivity in at least a relatively large proportion of the patientssuffering from one or other of the various forms of cancer which are orwill become accessible to the chemotherapy without, however, exceedingthose doses at which the substances would be too toxic.

As an example, the doses administered systemically, in particular byperfusion, and expressed as mg/kg, may vary from about 0.1 to about 5mg/kg, for example about 1 mg/kg.

The invention also relates to other forms of administration, inparticular by the oral route (liquid or solid compositions).

These dose ranges obviously have only indicative value.

It is of course to be understood that in this type of therapy the dosesadministered must be assessed by the clinician in each case, on whichoccasion the condition of the patient and his personal reactivitytowards medicines is taken into consideration.

An example of the pharmaceutical preparation of the products accordingto the invention contains from 10 to 250 mg, in particular 50 mg, of atleast one of the products according to the invention in the form--of asterile lyophilized powder in combination with an ampoule of aphysiologically acceptable solvent, in particular of physiologicalserum, containing 5 ml of the solvent per ampoule.

As a result of their particularly high activity, the substances of theinvention are also useful as reference compounds in pharmacologicalstudies, in particular in those designed to compare the antitumorproperties of substances under study in comparison with a referencecompound.

Example 33 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]2,3,6-trideoxy β-L-arabino-hexopyranoside (IC 85-1615) ##STR208## (a)Preparation of methyl 3-azido 2,3,6-trideoxy β-L-arabino-hexopyranoside.

3 g. (0.016 mole) of methyl 3-azido 2,3,6-trideoxy α-L-arabinohexopyranoside and 1 g. of toluene sulfonic acid in 50 ml of anhydrousmethanol are stirred for 48 hours at room temperature.

The reaction mixture is evaporated to dryness in a vacuum. The residueis taken up in methylene chloride. After being washed twice with water,the organic phase is dried over sodium sulfate, evaporated to dryness ina vacuum and chromatographed on a column of silica (eluant hexane-ethylacetate 5:1).

Analysis: C₇ H₁₃ N₃ O₃ =187.20

m.p.: 72°-73° (hexane)

[α]_(D) =+63.5° (c, 0.8 CHCl₃)

IR (Nujol) 3370 cm⁻¹ (OH) 2100 cm⁻¹ (azide)

(b) Preparation of methyl 3-amino 2,3,6-trideoxyβ-L-arabino-hexopyranoside.

This compound is prepared as indicated in example 8.

Analysis: C₇ H₁₅ NO₃ =161.20 Yield=85%

m.p.: 136°

[α]_(D) : +75.8° (c, 0.5 CHCl₃).

(c) Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]2,3,6-trideoxy β-L-arabino-hexopyranoside.

The procedure is the same as that indicated for example 9--in which2-chloroethyl isocyanate is used and nitrosation is carried out, forexample, with the aid of sodium nitrite to give the compound of theinvention, the physical properties of which are indicated below.

Analysis: C₁₀ H₁₈ ClN₃ O₅ =295.7

m.p.: 109°-110°

[α]_(D) =+22.8° (c, 0.5% CHCl₃)

Example 34 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]2,3,6-trideoxy β-L-arabino-hexopyranoside (IC. 85-1625) ##STR209##

This compound is prepared from methyl 3-amino 2,3,6-trideoxyβ-D-arabino-hexopyranoside as indicated, in particular, in thepreparation of the compound described in example 9--by using2-chloroethyl isocyanate and by nitrosating with aid, for example, ofsodium nitrite.

The physical properties of the compound of the invention are as follows:

Analysis: C₁₀ H₁₈ ClN₃ O₅ =295.7

m.p. 103°-105°

[α]_(D) =-25.0° (c, 0.3% CHCl₃)

Example 35 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]2,3-dideoxy β-D-arabino-hexopyranoside (IC 85-1673) ##STR210##

Preparation of methyl 3-azido 2,3-dideoxy β-D-arabino-hexopyranoside.

19.2 g. (0.095 mole) of methyl 3-azido 2,3-dideoxyα-D-arabino-hexopyranoside--and 1 g. of p-toluene sulfonic acid in 200ml of methanol are stirred for 48 hours at room temperature. The residueobtained after evaporation in a vacuum is taken up in 60 ml of anhydrouspyridine.

30 g. of acetic anhydride are added drop-wise while the temperature ismaintained at 15°. After being stirred for 16 hours the reaction mixtureis evaporated to dryness. The residue is taken up in methylene chloride;the organic phase is washed with a solution of 2N HCl, then with water,and finally with a solution of sodium bicarbonate. The organic phase isdried over sodium sulfate, evaporated in a vacuum and chromatographed onsilica using as eluant hexane: acetone 4:1 to give 19.5 g. of compound 3in the form of its diacetate and 2.5 g. of methyl 3-azido 2,3-dideoxyβ-D-arabino-hexopyranoside in the form of its diacetate.

This latter compound is taken up in 45 ml of anhydrous methanol and then5 ml of sodium methoxide are added. After being stirred for 4 hours thesolution is neutralized by the addition of Amberlite IRC50 resin. Afterevaporation the filtrate yields methyl 3-azido 2,3-dideoxyβ-D-arabino-hexopyranoside in the form of crystals.

Analysis: C₇ H₁₃ N₃ O₄ : 203.2.

Preparation of methyl-3-amino 2,3-dideoxy β-D-arabino-hexopyranoside.

This compound is prepared as indicated for example 8.

Analysis: C₇ H₁₅ NO₄ : 177

m.p.: 140°-142°

[α]_(D) : -61.8° (c, 0.55% MeOH).

Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido] 2,3-dideoxyβ-D-arabino-hexopyranoside.

The procedure is the same as that indicated, for example, for thepreparation of the compound described in example 9--in which2-chloroethyl isocyanate is used and nitrosation is carried out, forexample, by the aid of sodium nitrite to give the compound of theinvention with the following physical properties.

Analysis: C₁₀ H₁₈ ClN₃ O₆ : 311.72 Yield: 70%

m.p.: 68°-70°

[α]_(n) : -37.9° (c, 0.36% MeOH).

Example 36 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso-ureido]3,4,6-trideoxy α-D-xylo-hexopyranoside (IC 85-1590). ##STR211## (a)Preparation of methyl 2-O-acetyl 3-azido 4,6-dichloro 3,4,6-trideoxyα-D-galacto-hexopyranoside.

52 ml of sulfuryl chloride are added drop-wise to 44 g. (0.21 mole) of3-azido 3-deoxy D-gluco-pyranose in 160 ml of anhydrous pyridine and 200ml of chloroform cooled to -78° C. After being maintained for 2 hours at-78° C., the mixture is stirred for 5 hours at room temperature.

The reaction mixture is diluted with 400 ml of chloroform, then washedwith 2N hydrochloric acid, water, then with a solution of sodiumbicarbonate and again with water. After drying over sodium sulfate andevaporation to dryness in a vacuum, the residue is taken up in 200 ml ofmethanol to which is then added a 10% solution of potassium iodide in awater-methanol mixture (1:1).

After being neutralized by potassium bicarbonate, the solution isfiltered and evaporated and then the residue is taken up in chloroform.The organic phase is washed with a solution of sodium thiosulfate, thenwith water, dried over sodium sulfate and evaporated to dryness.

The residue is taken up in 100 ml of pyridine and the solution is cooledto 0°. 20 ml of acetic anhydride are added drop-wise at 0° C. and thesolution is stirred for one day, then the reaction mixture is evaporatedin a vacuum, the aqueous phase is extracted 2 or 3 times with 200 ml ofmethylene chloride. The organic phase is then washed with water,evaporated to dryness in a vacuum and 36 g. of a mixture correspondingto the 2 anomers α and β are obtained. The two anomers are isolated inthe pure state by chromatography on a column of silica (eluant:hexane-acetone 10:1).

α Anomer: 14.5 g.; m.p.: 74°-76° (hexane-ethyl acetate) [α]_(D) =+171°(c, 1.17 CHCl₃)

β anomer: 16.0 g.; m.p.: 108°-110° (hexane-ethyl acetate) [α]_(D) =-6,5°(c, 1.58 CHCl₃).

(b) Preparation of methyl 3-azido 4,6-dichloro 3,4,6,-trideoxyα-D-galacto-hexopyranoside.

5 g. (0.017 mole) of the preceding α-anomer are dissolved in 50 ml ofanhydrous methanol in the presence of 1 g. of p-toluene sulfonic acid.The reaction mixture is allowed to stand at room temperature for 18hours and then is evaporated to dryness in a vacuum. The residue istaken up in methylene chloride and the organic phase is washed twicewith water, dried, filtered and then evaporated.

The crystals obtained (4.2 g.) are recrystallized from a mixture ofhexane-ethyl acetate.

Analysis: C₇ H₁₁ Cl₂ N₃ O₃ : 256

m.p.: 139°-141°

[α]_(D) =+188° (c, 1.035 CH₃ OH).

(c) Preparation of methyl 3-amino 3,4,6-trideoxyα-D-xylo-hexopyranoside.

To 2 g. (0.0075 mole) of the preceding compound dissolved in 50 ml ofanhydrous toluene under nitrogen are added 0.5 g. of azo 2,2'bis-isobutyronitrile followed by 8 ml of tributyltin hydride addeddrop-wise. The reaction mixture is heated under reflux for 10 hours.After the mixture has been cooled and the precipitate has been removedfrom filtration, the filtrate is evaporated to dryness in a vacuum.Chromatography on silica (eluant: CH₂ Cl₂ -ammoniacal MeOH 9:1) leads tothe pure amino sugar in the form of white crystals.

Analysis: C₇ H₁₅ NO₃ : 161

m.p.: 136°-139° (ether-methanol)

[α]_(D) : +172° (c, 1% CHCl₃).

(d) Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-3-trideoxy α-D-xylo-hexopyranoside (IC 85-1591).

The procedure is the same as that for the preparation of the previouscompound.

Analysis: C₁₀ H₁₈ ClN₃ O₅ : 295.7 Yield: 65%

m.p.: 103°-105°

[α]_(D) : +138° (c, 1.41 CHCl₃).

Example 37

Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-trideoxy β-D-xylo-hexopyranoside (IC 85-1591). ##STR212##

(a) Preparation of methyl 2-O-acetyl 3-azido 4,6-dichloro 3,4,6-trideoxyβ-D-galacto-hexopyranoside.

52 ml of sulfuryl chloride are added drop-wise to 44 g. (0.21 mole) of3-azido 3-deoxy D-gluco-pyranose in 160 ml of anydrous pyridine and 200ml of chloroform cooled to -78° C. After being maintained for 2 hours at-78° C., the mixture is stirred for 5 hours at room temperature.

The reaction mixture is diluted with 400 ml of chloroform, then washedwith 2N hydrochloric acid, water, then with a solution of sodiumbicarbonate and again with water. After drying over sodium sulfate andevaporation to dryness in a vacuum, the residue is taken up in 200 ml ofmethanol to which is then added a 10% solution of potassium iodide in awater-methanol mixture (1:1).

After being neutralized by potassium bicarbonate, the solution isfiltered and evaporated and then the residue is taken up in chloroform.The organic phase is washed with a solution of sodium thiosulfate, thenwith water, dried over sodium sulfate and evaporated to dryness.

The residue it taken up in 100 ml of pyridine and the solution is cooledto 0° C. 20 ml of acetic anhydride are added drop-wise at 0° C. and thesolution is stirred for one day, then the reaction mixture is evaporatedin a vacuum, the aqueous phase is extracted 2 or 3 times with 200 ml ofmethylene chloride. The organic phase is then washed with water,evaporated to dryness in a vacuum and 36 g. of a mixture correspondingto the 2 anomers α and β are obtained. The two anomers are isolated inthe pure state by chromatography on a column of silica (eluant:hexane-acetone 10:1).

α Anomer: 14.5 g.; m.p.: 74°-76° (hexane-ethyl acetate) [α]_(D) =+171°(c, 1.17 CHCl₃)

β anomer: 16.0 g.; m.p.: 108°-110° (hexane-ethyl acetate) [α]_(D) =-6.5°(c, 1.58 CHCl₃).

(b) Preparation of methyl 3-azido 4,6-dichloro 3,4,6-trideoxyβ-D-galacto-hexopyranoside.

The procedure is the same as that described in part b of IC 85-1590.

Analysis: C₇ H₁₁ Cl₂ N₃ O₃ : 256 Yield: 85%

m.p.: 135°

[α]_(D) : +10.0° (c, 0.97% CH₃ OH).

(c) Preparation of methyl 3-amino 3,4,6-3-trideoxyβ-D-xylo-hexopyranoside.

The procedure is the same as that described in part c of IC 85-1590.

Analysis: C₇ H₁₅ NO₃ : 161

m.p.: 148°-149°

[α]_(D) : -52° (c, 1% CHCl₃).

(d) Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-trideoxy β-D-xylo-hexopyranoside.

The preparation procedure is the same as that for the precedingcompound.

Analysis: C₁₀ H₁₈ ClN₃ O₅ : 295.7 Yield: 90%

m.p.: 84°-90°

[α]_(D) : +11.5° (c, 1.25 CHCl₃).

Example 38 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-trideoxy α-L-xylo-hexopyranoside (IC 85-1626). ##STR213##

The compound is prepared according to the usual method starting frommethyl 3-amino 3,4,6-trideoxy α-L-xylo-hexopyranoside prepared accordingto H. Baer, Canad. J. Chem., vol. 52, 1974, p. 122-124.

Analysis: C₁₀ H₁₈ ClN₃ O₅ : 295.7 Yield: 65%

m.p.: 103°-105°

[α]_(D) =-141.5 (c, 0.71 CHCl₃).

Example 39 Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-trideoxy β-L-xylo-hexopyranoside (IC 85-1627). ##STR214##

The compound is prepared starting from methyl 3-nitro 3,4,6-trideoxyα-L-xylo-hexopyranoside described by H. Baer, Canad, J. Chem., 52, 1974,p. 122-124.

(1) Preparation of methyl 3-nitro 3,4,6-trideoxyβ-L-xylo-hexopyranoside.

40 ml of 1N hydrogen chloride in methanol are added to 1.8 g. of theabove α anomer. After being refluxed for 2 hours the reaction mixture isevaporated to dryness and the residue is purified on a column of silica(eluant: CH₂ Cl₂ -MeOH 98:2).

500 mg of the pure β anomer are obtained.

(2) Preparation of methyl 3-amino-3,4,6-trideoxyβ-L-xylo-hexopyranoside.

The preceding compound is dissolved in ethanol and hydrogenated atatmospheric pressure in the presence of platinum oxide.

Analysis: C₇ H₁₅ NO₃ : 161 Yield 80%

m.p.: 146°-148°

[α]_(D) =+36.0° (c, 0.96 CHCl₃).

(3) Preparation of methyl 3-[3-(2-chloroethyl) 3-nitroso ureido]3,4,6-trideoxy -L-xylo-hexopyranoside.

The procedure is the same as that previously described.

Analysis: C₁₀ H₈ ClN₃ O₅ : 295.7

m.p.: 90°-92° (isopropyl ether)

[α]_(D) : -9.0° (c, 0.6 CHCl₃)

PROTOCOL FOR TREATMENT USING THE PRODUCTS OF EXAMPLES 33-39

I. Materials and methods

A - Tumors used.

2 murine tumors were used for the "in vivo" studies: leukemia L1210 andmelanoma B16.

(1) LEUKEMIA L1210 Animals

The experiments were always carried out on female mice which werespecific pathogen-free (S. P. F.),

either of the line DBA/2JIco

or of the line B6 D2 F1/JIco (first generation hybrids between the linesC57BL/6 and DBA/2).

The line used in each experiment is specified.

Tumoral graft

On the day of the graft (by convention day 0=D0) an inoculum of 1×10⁵tumor cells in a volume of 0.2 ml is administered to each mouse by theintraperitoneal route (I.P.).

This inoculum is prepared by diluting ascites fluid taken from theperitoneum of a donor female mouse in NCTC 109 medium (EurobioLaboratories, Paris, France), counting of the cells in a MALASSEZ cellunder a microscope and adjustment of the concentration to 5×10⁵ cellsper liter by means of the same medium.

The tumor line is an American strain of the line L1210.

Distribution in experimental groups

After the tumor graft, the mice are distributed at random in cages of 5animals. Subsequently, these cages containing 5 mice are themselvesdistributed at random into a control group (control) and groups treatedwith the I.C. products.

(2) MELANOMA 16 Animals

The experiments are always carried out on SPF female mice, B6 D2 F1/JIco(first generation hybrids between the lines C57BL/6 and DBA/2).

Tumoral graft

On the day of the graft (day 0), an inoculum of 2×10⁶ tumor cells in avolume of 0.5 ml is administered to each mouse by the intraperitonealroute (I.P.). This inoculum is prepared from a sub-cutaneous tumorexcised from a donor female mouse. After excision the tumor isfragmented by means of a pair of scissors in the NCTC 109 medium. Afterfiltration through sterile gauze in order to remove large cellularfragments, the homogeneous cell suspension obtained is counted by meansof a MALASSEZ cell and diluted to the desired concentration (4×10⁶ tumorcells per ml) by dilution with the NCTC 109 medium.

Distribution in experimental groups

This was performed in the manner described for the leukemia L1210 (seeabove).

B - Protocol for treatment.

The doses of the products of the invention used in the differentexperiments are expressed in milligrams per kilogram of body weight.

The products to be injected were dissolved in isotonic sodium chloridesolution.

2 protocols for treatment were used in the different experiments:

either a single injection on D1, by the intraperitoneal (I.P.) orintravenous (I.V.) route,

or 3 I.P. injections on D1, D5, D9.

In each experiment the animals of the control group received 1 or moreinjections, depending on the experimental protocol used, by the sameroute (I.P. or I.V.) of the same volume (0.2 ml/20 g) of the vehiclewithout the active principle (isotonic sodium chloride solution).

II - Expression of results

For each experiment a table specifies:

the number and percentage of the total of the mice surviving to D60,

the T/C×100 for the treated groups.

T representing the mean survival time of the mice in the treated group,

C representing the mean survival time of the mice in the control group(controls).

III - Comment

The strains of mice used, the experimental protocols and the mode ofexpression of the results are in accordance with directive 271 F of the"N.C.I. Division of Cancer Treatment" (November 1983).

IV - Pharmacological results

IC. 1625

L1210 U.S.A. - DBA₂ - I.P. treatment on D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors at D60                                          ______________________________________                                        190       (0/10)                                                              ______________________________________                                    

IC. 1673

L1210 U.S.A. - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×1.25 mg/kg

    ______________________________________                                        T/C       Number of survivors                                                 ______________________________________                                        158       (0/10)                                                              ______________________________________                                    

L1210 U.S.A. - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×5 mg/kg

    ______________________________________                                        T/C       Number of survivors                                                 ______________________________________                                        626       (6/10)                                                              ______________________________________                                    

Melanoma B16 - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors                                                 ______________________________________                                        232       (4/10)                                                              ______________________________________                                    

IC. 1591

L1210 strain U.S.A.- DBA₂ - I.P. treatment on D1 1×20 mg/kg

    ______________________________________                                        T/C       Number of survivors on D60                                          ______________________________________                                        >600      (7/10)                                                              ______________________________________                                    

L1210 strain U.S.A. - DBA₂ - I.P. treatment on D1, D5, D9 3×5 mg/kg

    ______________________________________                                        T/C       Number of survivors on D60                                          ______________________________________                                        >600      (6/10)                                                              ______________________________________                                    

L1210 strain U.S.A. - B₆ D₂ F₁ - I.P. treatment on D1, D5, D9 3×1.25 mg,3×5 mg, 3×10 mg/kg

    ______________________________________                                                  T/C  Number of survivors on D60                                     ______________________________________                                        3 × 1.25                                                                            116    (0/10)                                                     3 × 5 217    (2/10)                                                     3 × 10                                                                              600    (8/10)                                                     ______________________________________                                    

Melanoma B16 - B₆ D₂ F₁ - I.P. treatment of D1, D5, D9 3×10 mg/kg

    ______________________________________                                        T/C       Number of survivors on D60                                          ______________________________________                                        194       (3/10)                                                              ______________________________________                                    

IC. 1626

L1210 strain U.S.A. - DBA₂ - I.P. treatment on D1 1×20 mg/kg

Number of survivors on D60: 3/10

L1210 strain U.S.A. - DBA₂ - I.P. treatment on D1, D5, D9 3×2.5 mg/kg,3×10 mg/kg

    ______________________________________                                                  T/C  Number of survivors on D60                                     ______________________________________                                        3 × 2.5                                                                             150    (0/10)                                                     3 × 10                                                                              275    (3/10)                                                     ______________________________________                                    

IC. 1627

L1210 strain U.S.A. - DBA₂ - I.P. treatment on D1, 1×20 mg/kg

Number of survivors on D60: 3/10

L1210 strain U.S.A. - DBA₂ - I.P. treatment on D1, D5, D9 3×2.5 mg/kg,3×10 mg/kg

    ______________________________________                                                 T/C   Number of survivors on D60                                     ______________________________________                                        3 × 2.5                                                                            141     (0/10)                                                     3 × 10                                                                             >600    (6/10)                                                     ______________________________________                                    

V - Toxicological results

DL0, DL50, single I.P. dose administered to the aforementioned strainsof mice.

For the compound in IC. 85-1625, the DL0 is higher than or equal to 40mg/kg.

For the compound in IC. 85-1590, the DL0 is higher than or equal to 25mg/kg.

For the compound in IC. 85-1591, the DL0 is higher than or equal to 20mg/kg.

For the compound in IC. 85-1626, the DL0 is higher than or equal to 20mg/kg.

For the compound in IC. 85-1627, the DL0 is higher than or equal to 40mg/kg.

For the compound in IC. 85-1673, the DL50 is approximately 20 mg/kg.

For the compounds in IC. 85-1591 and IC. 85-1626, the DL50 isapproximately 40 mg/kg.

We claim:
 1. A nitrosourea derivative of the formula: ##STR215## inwhich: R is hydrogen, alkyl of 1 to 30 carbon atoms, aralkyl of 7 to 12carbon atoms, or aralkyl of 7 to 12 carbon atoms substituted with Hal,--NO₂, NH₂, --CF₃ or alkoxy of 1 to 4 carbon atoms;X is hydroxy or --NR₁R₂ ; Y is hydrogen, hydroxy or --NR'₁ R'₂ ; R₁ and R'₁ are each hydrogenor ##STR216## Hal is a halogen; R₂ and R'₂ are each hydrogen, alkyl of 1to 6 carbon atoms, cycloalkyl of 3 to 6 carbon atoms, aryl of 4 to 10carbon atoms, aralkyl of 7 to 12 carbon atoms, aryl of 4 to 10 carbonatoms substituted with Hal, --NO₂, --NH₂, --CF₃ or alkoxy of 1 to 4carbon atoms, or aralkyl of 7 to 12 carbon atoms substituted with Hal,--NO₂, --NH₂, --CF₃ or alkoxy of 1 to 4 carbon atoms; R' and R" are eachhydrogen, hydroxy or --OM; and M is alkyl of 1 to 30 carbon atoms, arylof 4 to 10 carbon atoms, aralkyl of 7 to 12 carbon atoms, aryl of 4 to10 carbon atoms substituted with Hal, --NO₂, --NH₂, --CF₃ or alkoxy of 1to 4 carbon, aralkyl of 7 to 12 carbon atoms substituted with Hal,--NO₂, --NH₃, --CF₃ or alkoxy of 1 to 4 carbon atoms, acyl of 2 to 8carbon atoms, aroyl of 5 to 12 carbon atoms, or aroyl of 5 to 12 carbonatoms substituted with Hal, --NO₂, --NH₂, CF₃ or alkoxy of 1 to 4 carbonatoms; provided that at least:X is --NR₁ R₂ and R₁ is ##STR217## or Y is--NR'₁ R'₂ and R'₁ is ##STR218## and either R' is hydrogen or R" ishydrogen, and R' and R" are not both hydrogen.
 2. The nitrosoureaderivative of claim 1, wherein R is alkyl of 1 to 12 carbon atoms or asubstituted or unsubstituted aralkyl of 7 to 9 carbon atoms.
 3. Thenitrosourea derivative of claim 1, wherein Hal is Cl.
 4. The nitrosoureaderivative of claim 1, wherein R₂ and R'₂ are each substituted orunsubstituted aralkyl of 7 to 9 carbon atoms.
 5. The nitrosoureaderivative of claim 1, wherein M is alkyl of 1 to 12 carbon atoms,substituted or unsubstituted aralkyl of 7 to 9 carbon atoms, acyl of 2or 3 carbon atoms, or substituted or unsubstituted aroyl of 5 to 9carbon atoms.
 6. The nitrosourea derivative of claim 1, wherein: R' ishydrogen; and R" is hydroxy or --OM.
 7. The nitrosourea derivative ofclaim 1, wherein: R" is hydrogen and R' is hydroxy or --OM.
 8. Thenitrosourea derivative of claim 7, having the formula: ##STR219##
 9. Thenitrosourea derivative of claim 7, having the formula: ##STR220## 10.The nitrosourea derivative of claim 9, wherein R is alkyl of 1 to 12carbon atoms.
 11. The nitrosourea derivative of claim 9, wherein: R' ishydroxy.
 12. The nitrosourea derivative of claim 9, wherein Y ishydroxy.
 13. The nitrosourea derivative of claim 9, wherein Hal is Cl.14. The nitrosourea derivative of claim 13, wherein: R is alkyl of 1 to12 carbon atoms; R' is hydroxy; and Y is hydroxy.
 15. The nitrosoureaderivative of claim 9, wherein: Y is --NR'₁ R'₂ ; and R'₁ is ##STR221##16. The nitrosourea derivative of claim 15, wherein: R is alkyl of 1 to12 carbon atoms or aralkyl; and R' is hydroxy.
 17. The nitrosoureaderivative of claim 16, wherein: X is --NR₁ R₂ ; and R₁ is ##STR222##18. The nitrosourea derivative of claim 1, wherein: R is alkyl of 1 to12 carbon atoms or aralkyl; R' or R" is --OM; M is alkyl of 1 to 12carbon atoms or aryl; X is --NR₁ R₂ ; R₁ is ##STR223## and Y is hydrogenor hydroxy.
 19. The nitrosourea derivative of claim 1, wherein: R isalkyl of 1 to 12 carbons atoms or aralkyl; R' or R" is hydroxy; X is--NR₁ R₂ ; R₁ is ##STR224## and Y is hydrogen.
 20. The nitrosoureaderivative of claim 1, wherein: R is alkyl of 1 to 12 carbons atoms oraralkyl; X is hydroxy; Y is --NR'₁ R'₂ ; and R'₁ is ##STR225##
 21. Thenitrosourea derivative of claim 20, wherein: R' or R" is hydroxy. 22.The nitrosourea derivative of claim 1, selected from the group ofcompounds having the formula: ##STR226##
 23. A nitrosourea derivative ofthe formula: ##STR227## in the form of one of the two anomers, α or β,in which: R₁ is alkyl of 1 to 12 carbon atoms, aralkyl of 7 to 12 carbonatoms or aralkyl of 7 to 12 carbon atoms substituted on the aromaticnucleus with Hal, --NO₂, --CF₃ or alkoxy of 1 to 4 carbon atoms;R₄ is Hor Hal; Hal is a halogen; R₆ is --OH or ##STR228## R is alkyl of 1 to 6carbon atoms, aryl, aryl substituted on the aromatic nucleus with Hal,--NO₂, --CF₃ or alkoxy of 1 to 4 carbon atoms; and Nu is ##STR229## 24.The nitrosourea derivative of claim 23, wherein R₁ is aralkyl orsubstituted aralkyl of 7 to 9 carbon atoms.
 25. The nitrosoureaderivative of claim 23, wherein Hal is chlorine.
 26. The nitrosoureaderivative of claim 23, wherein: R₁ is alkyl of 1 to 12 carbon atoms;and R₄ is Hal.
 27. The nitrosourea derivative of claim 23, wherein R₁ is--CH₃.
 28. The nitrosourea derivative of claim 27, wherein R₄ ischlorine.
 29. The nitrosourea derivative of claim 27, wherein R₆ is##STR230##
 30. The nitrosourea derivative of claim 23, wherein: R₁ is--CH₃ ; R₄ is H; and R₆ is --OH.
 31. The nitrosourea derivative of claim23, wherein: R₁ is --CH₃ ; R₄ is H; and R₆ is ##STR231##
 32. Thenitrosourea derivative of claim 23, selected from the group of compoundshaving the formula: ##STR232##
 33. A method for treating a patient witha tumor, comprising the step of administering to the patient aneffective anti-tumor amount of a nitrosourea derivative of any one ofclaims 1 to
 32. 34. The method of claim 33, wherein the nitrosoureaderivative is administered to the patient in a dosage of about 1 to 50mg per kg of the patient.
 35. The method of claim 33, wherein thenitrosourea derivative is administered orally to the patient.
 36. Apharmaceutical composition for treating a patient with a tumor,comprising: an effective amount of a nitrosourea derivative of any oneof claims 1 to 32; and a pharmaceutically acceptable vehicle.